CZ2001241A3 - Phthaazine derivatives - Google Patents

Abstract

The phthalazine derivative of formula (I); Z is NH, methylene, an alkylene chain which is optionally branched and / or unsaturated and / or interrupted by a (C5-C7) cycloalkyl residue; A is phenyl or heterocycle optionally substituted with one or more substituents selected from oxo, nitro, carboxy and halogen, or COR 4; R represents an alkyl or polyfluoroalkyl group; R 1 is absent when it is a double bond, or when a single bond represents a) a hydrogen atom; b) alkyl optionally substituted with aryl, heterocycle or COR 5; c) -CORO; d) alkylsulfonyl; R 2 represents two hydrogen atoms or = O, wherein -— is a single bond, or if -I is a double bond, R 2 is hydrogen, cyano, alkoxycarbonyl, amido, optionally substituted aryl or heterocycle, alkyl, alkenyl or alkynyl or branched and / or substituted; R 3 is hydrogen, or and alkyl, alkenyl or alkynyl optionally substituted with hydroxy, oxo, aryl or heterocycle, and optionally interrupted by one or more heteroatoms or one or more hetero groups; N? Derivatives of the compounds of formula I and pharmaceutically acceptable salts thereof. Also described are pharmaceutical compositions containing these compounds and the use of compounds as PDE 4 inhibitors.

Description

Phthalazine derivatives

Technical field

The present invention relates to phthalazine derivatives, pharmaceutical compositions containing them and their use as phosphodiesterase 4 inhibitors.

BACKGROUND OF THE INVENTION

Phosphodiesterases belong to the group of isoenzymes that form the main mechanism of hydrolytic inactivation of cAMP (cyclic adenosine-3 ', 5'-monophosphate). CAMP has been shown to be the second messenger mediating a biological response to many hormones, neurotransmitters, and drugs (Krebs Endocrinology Proceedings of the 4th International Congress Excerpt Medica 17, 1973). When a suitable agonist binds to the cell surface, adenylate cyclase activates and converts Mg ²⁺ -ATP to cAMP. cAMP modulates the activity of most, if not all, cells contributing to the pathophysiology of various respiratory diseases, both allergic and of other origin. Accordingly, increasing cAMP levels will provide beneficial effects such as airway smooth muscle relaxation, inhibition of stem cell mediator release (basophilic granulosa cells), inhibition of neutrophil and basophil degranulation, inhibition of monocyte and macrophage activation. Thus, compounds capable of activating adenylate cyclase or inhibiting phosphodiesterases could suppress unwanted airway smooth muscle activation and a large number of inflammatory diseases.

Phosphodiesterases include a class of isoenzymes, phosphodiesterases 4 (PDE 4), specific for cAMP hydrolysis in airway smooth muscle and inflammatory cells (Torphy,

Phosphodiesterase Isoenzymes: Potential Targets for Novel Antiasthmatic Agents in New Drugs for Asthma, Barnes, ed. IBC Technical Services Ltd. 1989). Studies conducted on this enzyme indicate that its inhibition not only leads to airway smooth muscle realization, but also inhibits degranulation of mast cells, basophils and neutrophils, as well as inhibition of monocyte and neutrophil activation. PDE 4 inhibitors are thus effective in the treatment of asthma. These compounds offer a unique approach to the treatment of various respiratory diseases, both allergic and non-allergic, and have substantial therapeutic advantages over conventional therapy.

The excessive or irregular production of tumor necrosis factor (hereinafter _TNF), a cytokine with pro-inflammatory activity produced by various kinds of cells, affects the outbreak of many pathologies such as adult respiratory distress syndrome (adult respiratory distress syndrome, ARDS) and the chronic pulmonary inflammatory diseases. Therefore compounds able to reduce the negative effects of TNF _a, ie. Inhibitors of this cytokine, considered useful for the treatment of many pathologies.

Patent application EP 722 936 (Eisai) claim inter alia a compound of the formula

where n = 0-4; R 6 is an optionally substituted lower alkoxy group, an optionally substituted cycloalkyl group, or a -OR 8 group wherein R 8 is an optionally substituted arylalkyl group; X is -N = or -NR 8 -, wherein R 6 is hydrogen, lower alkyl, or optionally substituted arylalkyl, or

heteroarylalkyl; Y is -CO or -CB = wherein B is -NR ₇ R ₈ , where one of R ₇ and R 8 may be hydrogen and the other is optionally substituted heteroaralkyl, or B is hydrogen or optionally substituted aryl, heteroaryl, arylalkyl or heteroarylalkyl; A is hydrogen or halogen, or an optionally mono- or disubstituted amino group, optionally substituted aryl, heteroaryl or heteroarylalkyl groups. Among the groups which may optionally substitute the abovementioned radicals are halogen atoms. These compounds are intended to be effective as inhibitors of cGMP-PDE, i.e. PDE 5, a phosphodiesterase acting by the cGMP-dependent mechanism, whose field of application is markedly cardiovascular (Schudt C. et al., Phosphodiesterase Inhibitors, Academic Press).

Patent application EP 634 404 (Rhone Pouleno Agriculture) describes, inter alia, phthalazinones of the formula

wherein R represents an arylalkyl group, especially pyridyl optionally substituted with halogen atoms; R ₁ represents an alkyl chain containing up to 6 carbon atoms or an arylalkyl group, especially phenyl; R2 is phenoxy or benzyloxy; and m = 0-4. These compounds are useful as pesticides.

U.S. Pat. No. 3,274,185 to Messengill discloses, inter alia, phthalazines of the formula

OF

Wherein Y and Y 1 are independently hydrogen or lower alkoxy; Z is phenyl optionally substituted with halogen, or benzyl optionally substituted with lower alkyl or alkoxy; and R is hydrogen. Phthalazines have sedative and hypotensive effects, with no mechanism of action.

U.S. Pat. No. 3,813,384 to Asta-Werke illustrates, inter alia, benzylphthalazinones of the formula

wherein R 1 and R ₂ are hydrogen, lower alkoxy or halogen; X is an optionally branched alkylene chain; m and n are 13; p is 0 or 1; and the -cQr group is optionally substituted with a (C3-C8) mono-, di- or tricyclic radical containing one or two nitrogen atoms. These compounds have histaminolytic effects and are useful, for example, in the treatment of asthma. All compounds exemplified contain a residue which is a saturated heterocycle.

WO 97/40020 (Schering AG) discloses compounds of the formula

• · • ·

wherein Y is -NR ₃ - or -N =, and R ₂ is H, lower alkyl, nitro, halogen, amino, lower alkoxy or -CF ₃ , R ₃ is H, -CO- substituted H, lower alkyl substituted with aryl , amino, lower alkoxy, cycloalkyl or cycloalkoxy, or R ₃ is lower alkyl or cycloalkyl, R ₄ is H or lower alkoxy, R 5 is lower alkyl. These compounds are said to be non-competitive antagonists of excitatory amino acids.

Patent application WO 97/48697 (Rhone Poulenc Rorer) describes bicyclic compounds with inhibitory effects on TNF and PDE 4 _and that are represented by a very broad general formula. The phthalazine compounds could be included in the general formula of this patent application, however, none of the compounds disclosed in the patent examples is a phthalazine derivative and this kind of structure is excluded from the claims.

Patent application EP 848 000 (Tanabe Seiyaku) describes, inter alia, phthalazine derivatives of the formula

wherein R 1 and R ₂ are H, or an optionally protected hydroxy group: R ₃₃ is lower alkyl; R 5 and R 6 are H, amino or can form a heterocycle. These compounds are PDE 4 inhibitors.

It has now been surprisingly discovered that a new class of phthalazine derivatives able to inhibit PDE 4 and _TNF.

SUMMARY OF THE INVENTION

The present invention therefore relates to compounds of formula I

σ) where - represents a single or double bond;

Z is NH, methylene, (C ₂ -C ₆₎ alkylene chain which is optionally branched and / or unsaturated and / or interrupted by a (C5-C7) cycloalkyl radical;

A is phenyl or heterocycle optionally substituted with one or more substituents selected from oxo, nitro, carboxy and halogen, or COR ₄ wherein R ₄ is hydroxy, (C 1 -C ₆ ) alkoxy, amino optionally substituted with one or two (C 1 -C ₆ ) -C ₆ ) alkyl or hydroxy;

R is (C 1 -C 6) alkyl or polyfluoro (C 1 -C 6) alkyl;

R 1 is absent when Z 2 is a double bond or when Z 2 is a single bond means

(a) a hydrogen atom;

b) (C 1 -C 6) alkyl optionally substituted with aryl, heterocycle or COR ₅ wherein R 5 is hydroxy, (C 1 -C 4) alkoxy or hydroxyamino;

c) -COR ₆ wherein R ₆ is hydrogen, aryl, aryl-C 1 -C 6 alkyl, amino optionally alkylated or monohydroxylated, hydroxy, (C 1 -C ₄ alkoxy, carboxy, (C 1 -C ₄ ) alkoxycarbonyl, HN = C-NH ₂ , or (C 1 -C 4) alkyl optionally substituted with heterocycle;

d) (C 1 -C 4) alkylsulfonyl;

R ₂ represents two hydrogen atoms or the group = 0, where 22η is a single bond, or when - is a double bond, R ₂ is hydrogen, cyano, (Ci-C4) alkoxycarbonyl, amido, optionally substituted aryl or heterocycle, (C C ₁ -C ₈ alkyl, (C ₂ -C ₈ ) alkenyl or (C ₂ -C ₈ ) alkynyl optionally branched and / or substituted with aryl or heterocycle; aryloxy, heterocyclyloxy, aryl- (C 1 -C 4) -alkoxy, heterocyclyl- (C 1 -C 4) alkoxy, amino substituted with one or two (C 1 -C ₄ ) alkyl groups, aryiamino, heterocyclylamino, aryl- (C ₁ -C 4) alkylamino , heterocyclyl- (C 1 -C 4) alkylamino;

R ₃ is hydrogen, or a (C ₁ -C ₈ ) alkyl, (C ₂ -C ₈ ) alkenyl or (C ₂ -C ₈ ) alkynyl optionally substituted with hydroxy, oxo, aryl or heterocycle, and optionally interrupted by one or more heteroatoms or one or more hetero groups;

N -> O derivatives of compounds of formula I and their pharmaceutically acceptable salts.

Preferred compounds of the present invention are compounds of formula I wherein 2122 represents a double or single bond; and Z is methylene or a _(C2-C8) alkylene chain. Even more preferred compounds of the invention are compounds of formula I wherein - is a double or single bond; Z is methylene or a C ₂ -C ₆ ) alkylene chain; and A represents a heterocycle optionally substituted by one or more; and A represents a heterocycle, optionally substituted by one or more. substituents; even more preferred compounds of the invention are compounds of formula I wherein - is a double or single bond; Z is methylene; and A is pyridine substituted with two substituents.

Within this class, a preferred group of compounds is represented by compounds of formula IA

(I-A) (of formula I wherein Z is methylene and A is 3,5-dichloropyridin-4-yl).

More preferred compounds are compounds of formula Ia, where 2222 is a single bond and _R2 represents two hydrogen atoms.

Another class of even more preferred compounds are compounds IA wherein 222: represents a double bond; and R ₂ is cyano, _(Ci-C4) alkoxycarbonyl, amido, optionally substituted heterocycle, _(C2-C8) alkenyl or (C ₂ -C) alkynyl optionally substituted by aryl or heterocycle; aryloxy, heterocyclyloxy, arylamino, heterocyclylamino.

In this class of compounds, compounds of formula IB are particularly preferred

(I-B) • 000 · · 0 · 0 0 · 0 0 0000 00 0 0 0 0 0 0 0000 0 0 0 0

000 000 000 00 0000 00 0 00 000 (of formula 1A, where 2 ^ 2 is a double bond, R ₃ is hydrogen, R is methyl), wherein R ₂ is a heterocycle.

Specific examples of preferred compounds of the invention are the following:

1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-4-phenyl-phthalazine;

4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazine-2-carboxylic acid methyl ester;

benzyl- {3- [1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl] -prop-2-ynyl} -methylamine;

1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-5- (5-morpholin-4-yl-pent-1-ynyl) -phthalazine dihydrochloride;

3- [1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl] -prop-2-yn-1-ol;

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4-morpholin-4-yl-phthalazine;

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4- [1,2,4] triazol-1-yl-phthalazine.

The compounds of formula I may contain one or more asymmetric centers and may therefore be in the form of stereoisomers. The present invention provides compounds of formula I both in the form of stereoisomeric mixtures and in the form of individual stereoisomers.

Compounds of formula I are active as PDE 4 and _TNF, and thus are used as therapeutic agents in allergic and inflammatory pathologies such as COPD, asthma and allergic rhinitis.

Heterocycles include, in particular, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, piperazine, triazole, morpholine, pyrrolidine, pyrroline, imidazoline, pyrazoline, pyrazolidine,

Imidazolidine, piperidine, furan, pyran, thiazole, isothiazole, isoxazole, thiophene and the like.

Halogen means fluorine, chlorine, bromine or iodine.

Particular examples of alkyl groups are methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, tert-butyl, n-pentyl, 1-methyl-butyl, 2-methylbutyl, 3-methylbutyl, 3- methyl (2-butyl, n-hexyl, heptyl, octyl and the like) The group (C 5 -C 8 cycloalkyl means cyclopentyl, cyclohexyl and cycloheptyl, while aryl is an aromatic ring or a system of 6 to 10 carbon atoms, with aryl and aryl- (C 1 -C 6) alkyl include phenyl, benzyl, phenethyl, phenylpentyl, naphthyl, indanyl, indanylpentyl and the like.

The oxidized form of N - O, if present, may include both the nitrogen atoms present on the phthalazine ring as well as the nitrogen atoms present on the group A when the group is a heterocyclic substituent.

Pharmaceutically acceptable salts of the compounds of formula I are salts with organic and inorganic acids, such as hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, acetic, benzoic, maleic, fumaric, succinic, tartaric, citric, aspartic, methanesulfonic, 3, 7-di-tert-butylnaphthalene-1,5-disulfonic acid (dibudinic acid) or with inorganic bases such as sodium or potassium hydroxide or sodium bicarbonate.

The synthesis of compounds of formula I proceeds by methods known to those skilled in the art. For example, starting from a compound of formula II

4 »999

. 4 9 4 4 9

9444999 9 · 4 4 9 9 9 4 4

4499 99 9 99 494

Wherein R 1 and R 3 are as defined above, and R 2a is a hydrogen atom, an optionally substituted aryl or heterocycle, or (C 1 -C 8) alkyl, (C 2 -C ₈ ) alkenyl or (C 2 -C ₈ ) alkynyl optionally substituted with an aryl or heterocycle group which For example, treatment of a compound of formula III

wherein R and R 3 are as defined above, treatment with a strong base, for example n-butyllithium, provides a compound of formula IIIa

(IDa) wherein R and R 3 are as defined above and M is lithium or sodium which, by treatment with a formyl electrophilic reagent, provides a compound of formula II, R 2a is a hydrogen atom. This compound can be converted to a compound of formula II wherein R 2a is different from hydrogen by treatment with a suitable organometallic reagent such as a Grignard reagent to give a compound of formula IV

- 12 ·· ·· β ·· · · · · · · · · · ·

4 9 9 9 4 4 4 4 * · ··· ··· «··« 4

Wherein R, R _2a and R ₃ are as defined above, wherein this compound provides oxidation, for example with pyridine-SO 3, DMSO and triethylamine, to provide the desired compound II.

A compound of formula IV can also be obtained from a compound of formula IIIa by reaction with an aldehyde of formula V

Raa-CHO (V), wherein R _2a is as defined above, used in molar excess relative to compound IIIa. A compound of formula II wherein R _2a is different from a hydrogen atom may also be synthesized directly from a compound of formula IIIa by transmetallation with a suitable salt, for example zinc chloride, followed by reduction with acyl chloride in the presence of a transition metal catalyst, for example palladium.

After cleavage of the oxazoline group from the compound of formula II, reaction with hydrazine in a protic solvent is carried out to give the phthalazinone of formula VI

wherein R, R _2a and R ₃ are as defined above which, by treatment with a halogenating agent such as phosphorus pentoxide, provides a compound of formula VII

(vn)

X • φ • φ φφφφ

wherein R, R _2a and R 3 are as defined above, and X represents a bromine or chlorine atom. This material is treated with a compound of formula VIIIa

AZY (VIII) wherein A and Z are as defined above and Y is a metal salt, for example Li, Na, Mg or a transition metal complex, a compound of formula I wherein R ₂ has the meanings given above when J is ^a double bond and R ₄ is not present.

If a compound of formula I wherein R 1 is present, compounds of formula I wherein - is a double bond and R ₂ is hydrogen, is to be prepared by known methods. For example, reduction with hydrogen in the presence of Pd / C or PtO ₂ yields a compound of formula I wherein R 1 is a hydrogen atom, followed by treatment with a suitable sulfonating or acylating agent to give a compound of formula I wherein R 1 is (C 1 -C 4) alkylsulfonyl or -CO-R ₆ , wherein R 6 is as defined above.

Alternatively, compounds of formula I may be obtained from an acid of formula VIIIa

R '

COOH where R and R3 are as defined above, which, by reaction with formaldehyde and HCl, provides a compound of formula IX

R ·

99 · · · 9 9 9

9 9 9 9 9

9 9 99 9 9999

9 9 9 9 9 9

9

14 where R and R3 are as defined above. This compound is halogenated, for example with N-bromosuccinimide, in the presence of a catalyst such as benzoyl peroxide, or 2,2'-azoisobutyronitrile, to give

wherein R, R 3 and X are as defined above treated with triphenylphosphine to provide a compound of formula XI

wherein R, R 3 and X are as defined above that react with an aldehyde of formula XII

A-Z'-CHO (XII) wherein A is as defined above and Z 'is methylene or a (C ₂ -C ₅ ) alkylene chain optionally branched and / or unsaturated and / or interrupted by a (C ₅ -C ₇ ) cycloalkyl radical or is absent, in the presence of an organic base such as triethylamine, to give a compound of formula XIII (ΧΙΠ)

Z · —A · * «· 44 4

4 9 4 4 4 4 4 44

4 4 4 4 4 4 4 4 4 • 4 4 444 4949 494 · ··· 9 4 4 4 4 4

IV · 9 ···· · 9 9 44 444 where R, R ₃ , Z 'and A are as defined above. This compound is reacted with hydrazine to give a compound of formula I wherein R 1 is hydrogen, R ₂ is = O, and Z is methylene, or a (C ₂ -C ₅ ) alkylene chain optionally branched and / or unsaturated and / or interrupted (C5- _C7 ) cycloalkyl radical, but not on the first carbon atom. Reaction of this compound of formula I with a suitable alkyl halide or sulfonate in the presence of a base such as sodium hydride provides a compound of formula I wherein R 1 is a substituent other than hydrogen.

Compound of Formula I wherein R 1 is a hydrogen atom, may also provide other compounds of formula I wherein Z is methylene or a (C ₂ -C 8) alkylene chain optionally branched and / or unsaturated and / or interrupted by a (C ₅ -C ₇ ) cycloalkyl radical, but not on the first carbon atom . For example, this compound is reacted with a halogenating agent such as POCl ₃ or POBr ₃ to give an intermediate of formula XIV

wherein A, R, R ₃ , Z and X are as defined above, which is then subjected to a coupling reaction in the presence of a catalyst such as palladium, or an aromatic nucleophilic substitution, to give a compound of formula I wherein R ₂ has the meanings given above - is a double bond, and Z is a methylene or (C ₂ -C ₅ ) alkylene chain optionally branched and / or unsaturated and / or interrupted by a (C ₅ -C ₇ ) cycloalkyl radical, but not on the first carbon atom.

As regards the substituent R _3, if it is other than a hydrogen atom, it may already be present in the starting products of the various ones described above. 9 99 99 9 99 9

9999 999 9 9 99 • 9 9 9999 999

. 9 9 99 9999999 9 9

-. 1 fi - ··· 99 9 99 9

IV 99 9999 99 9 99 99 synthetic routes, or may be introduced and modified in the process by methods known to those skilled in the art. For example, if R ₃ is (C 2 -C 6) alkenyl or (C 2 -C ₈ ) alkynyl optionally substituted with aryl or heterocycle, it may be hydrogenated to provide the corresponding (C 1 -C 6) alkyl or (C ₂ -C 8) alkenyl radical. methods known to those skilled in the art.

Alternatively, with a compound of formula XV

(XV) wherein R 1 and R 2 are as defined above, activates on a hydroxyl group, for example trifluoromethanesulfonic anhydride, to give a compound of formula XVI

(XVI) wherein R and R2 are as defined above and W represents an activating group. This compound is then treated with a coupling reaction in the presence of a catalyst such as palladium to give the desired compound of formula VI, which is then treated as above to give a compound of formula I.

The compound of formula XV can be obtained, for example, from a compound of formula XVII

R '

- 18 44 «4 · 4 4 • 4 4

4 4

4 4 444

4 4

4 4 4 4

Bz — O ^{Z CHO} (ΧΥΠ) wherein R is as defined above and Bz represents a benzyl group that is oxidized, for example with potassium permanganate and tetrabutylammonium bromide to give an acid of formula XVIII .O

Bz — O

R '

COOH (xvm) wherein R and Bz are as defined above, which is converted, for example, with thionyl chloride to the corresponding acyl halide of formula XIX, O

Bz-0

COX (XIX) wherein R, Bz and X are as defined above. This compound is reacted with diethylamine in at least equimolar amounts to give the benzamide of formula XX

Bz-0 _from Et

CO— (XX)

Et

- 18 • »99

9 9 9

9 9

9 9

9 9

9999

9

9

9999999 9

Wherein R and Bz are as defined above, which reacts with dimethylformamide in the presence of a strong organic base such as butyllithium, tert-butyllithium, sec-butyllithium, optionally in the presence of ligands such as tetramethylethylenediamine, to provide a compound of formula XXIa

(XXIa) wherein R 1 and B 2 are as defined above and R 2 'is a hydrogen atom. This compound is reacted with hydrazine in acetic acid to give a compound of formula XXII

wherein R, Bz and R2 'are as defined above which are debenzylated with HCl in acetic acid to provide a compound of formula XV.

Compound XXII, treated by methods already described for analogous compounds (e.g., compound XVI) can also provide compound XXIII

XX (XXIII) · 99 · 9 9

99 · 9,999,9999 <9,999,999 9 9

9 9 99 99 * 9999 9 '9

...

9999 99 99 99 999

Wherein R, R ₂ ', W, Z and A are as defined above which are reacted under conditions allowing the formation of a bond in the presence of palladium to give a compound of formula I wherein R ₂ is hydrogen.

An alternative to obtain a compound of formula I wherein R ₂ is different from a hydrogen atom is believed to be treated with a compound of formula XXIa with a R ₂ "-magnesium halide, for example chloride, or a R ₂ " -lithium compound wherein R ₂ "is R ₂ above, except for hydrogen, to obtain a compound of formula XXIV

(XXIV) wherein R, Bz and _R2 "are as defined above. The compound of formula XXIV is treated with a suitable oxidizing agent such as pyridinium chlorochromate to give a compound of formula XXIb

wherein R and R ₂ "are as defined above. Compounds of formula XXIa or XXIb are treated with acetic acid in an acidic medium to provide a compound of formula XXV

O (XXV)

- 20 · 9 • 9 • β β β β β β β β β 20 20 20 20 20 20 20 20 20 Wherein R and R 2 are as defined above that react with hydrazine to provide phthalazinone of formula XV.

The synthesis of the N-oxide of the compounds of formula I is carried out by reacting the compounds of formula I with peracids, such as, for example, chloroperbenzic acid.

The preparation of the salts of the compounds of formula I is carried out by known methods.

The compounds of formula I are PDE 4 inhibitors, as shown by in vitro enzyme inhibitory activity assays (Example 147), and are also capable of inhibiting TNFα release (Example 148). Comparisons were made with the following compounds: 6,7-dimethoxy-4- (pyridin-4-ylmethyl) -2 H -phthalazin-1-one (ref. 1) and 6,7-dimethoxy-4- (piperidin-4). -ylmethyl) -2H-phthalazin-1-one (ref. 2) included in the general formula already cited in patent application EP 722 936 (Eisai) and selected in terms of structural affinity with the compounds of the present invention. Reference compounds, although chemically similar, did not show effects as PDE 4 inhibitors.

In addition, the compounds of the present invention have no effects on PDEs 3 and 5 (Example 149).

Obviously, this selectivity and specificity in conjunction with the absence of an effect on the cardiovascular system make the compounds of Formula I useful for treating pathologies in which PDE 4 and TNF are involved _and , such as asthma, chronic obstructive pulmonary disease (COPD), respiratory insufficiency syndrome. adult (ARDS), allergic rhinoconjunctivitis, psoriasis, atopic dermatitis, rheumatoid arthritis, septic shock, ulcerative colitis, although the context of the present invention is particularly directed to pathological conditions of the respiratory tract. The compounds of the present invention are particularly useful in the treatment of allergic and inflammatory diseases, and in particular in the treatment of COPD, asthma and allergic rhinitis.

- 21 ΦΦ · · 9 * * * * * * * * * * 4 4 4

·· · · · · ·

9 9 ·· ···

Therapeutic doses will generally be from 0.1 to 1000 mg per day and from 1 to 200 mg per single dose for oral administration.

A further object of the present invention are pharmaceutical compositions comprising a therapeutically effective amount of the compounds of formula I or pharmaceutically acceptable salts thereof in admixture with a suitable carrier.

The pharmaceutical compositions of the present invention may be liquid, suitable for enteral or parenteral administration, and preferably solid such as tablets, capsules and granules suitable for oral administration, or may be in a form suitable for transdermal and inhalation administration.

The preparation of the pharmaceutical compositions of the present invention can be accomplished by known methods.

The following examples are provided to better illustrate the invention.

¹ H-NMR spectra were measured at 200 MHz on a Varian instrument; δ are in parts per million.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

2- (4-Methoxyphenyl) _{-4,4-dimethyl-4-one} 5 dihydrooxazol

To a solution of 2-amino-2-methyl-propan-1-ol (10.45 g, 1.17 mol) in CH 2 Cl 2 (400 mL) under N ₂ was added dropwise a solution of 4-methoxybenzoyl chloride (100 mL) over 30 min. g, 0.59 mol) in CH ₂ Cl ₂ (500 mL), maintaining the temperature at 18 ° C with water / ice. After stirring for 3 hours, the precipitate was filtered through celite and washed with CH ₂ Cl ₂ . The organic phase was stirred under N ₂ at 2 ° C and thionyl chloride (123 mL, 1.77 mol) was added dropwise, keeping the temperature below 10 ° C. Drip after completion! The reaction mixture was stirred overnight, then concentrated in vacuo. The residue was taken up in 5% HCl and twice

- 22 · φ · φ * * * φ * φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ Extracted with ethyl ether. The extracts were extracted with 5% HCl. The aqueous phase was basified with concentrated NaOH and extracted three times with ethyl ether, then dewatered and concentrated in vacuo. The crude was distilled at 95 to 98 ° C (30 Pa) to give 106.43 g of the title compound (yield: 87.6%).

¹ H-NMR (CDCl 3) δ: 7.87-6.84 (m, 4H); 4.04 (s, 3H); 3.80 (s, 3H); 1.34 (s, 6H).

Example 2

2- (4,4-Dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxybenzaldehyde

2- (4-Methoxyphenyl) -4,4-dimethyl-4,5-dihydrooxazole (20 g, 0.097 mol) prepared as described in Example 1 was dissolved in ethyl ether (200 mL) and dried under N ₂ atmosphere. n-butyllithium (44 mL, 0.11 mol) was added at -2 ° C while maintaining the temperature of the mixture below 5 ° C. The mixture was stirred and after 4 h DMF (15.4 g, 16.3 mL, 0.21 mol) was added. After stirring overnight at room temperature, the mixture was cooled and extracted with water / ice. The aqueous phase was extracted again with ethyl ether. The organic phase was washed with water / NaCl, decolourised with charcoal, dewatered and concentrated in vacuo to give 19 g of the title compound (yield: 84%).

¹ H-NMR (CDCl 3) δ: 10.75 (s, 1H); 7.84-7.05 (m, 3H); 4.10 (s. 2H); 3.86 (s, 3H); 1.37 (s, 6H).

Example 3

6-Methoxy-2H-phthalazin-1-one

A solution of 2- (4,4-dimethyl-4,5-dihydro-oxazol-2-yl) -5-methoxybenzaldehyde (2 g, 8.58 mmol), prepared as described in Example 2, in 30 mL of a mixture water (5 mL), ethanol (50 mL) and concentrated H ₂ SO ₄ (4 mL) made up to 100 mL with ethanol, stirred at reflux for 20 h, then concentrated to small volume, taken up in water, extracted with ethyl ether and organic phase was washed with water, dewatered and dried. The obtained oil was dissolved in acetic acid (14 mL) and hydrazine monohydrate (1.25 mL, 25.7 mmol) dissolved in acetic acid (6 mL) was added dropwise under N ₂ atmosphere. The mixture was stirred at room temperature for 3.5 hours, then at 80 ° C for 4 hours, cooled, almost completely dried, taken up in water, neutralized with NaHCO ₃ , extracted several times with CH ₂ Cl ₂ . The organic phases were washed with water, dewatered and concentrated. The residue was triturated in CH ₂ Cl ₂ (10 mL) and filtered to give 1.1 g of the title compound (yield: 73.4%). Melting point: 200 - 205 ° C.

¹ H-NMR (DMSO) δ: 12.50 (s-broad, 1H); 8.26 (s, 1 H); 8.14-7.37 (m, 3H); 3.91 (s, 3H).

Example 4

1-Chloro-6-methoxyphthalazine

To a suspension of 6-methoxy-2H-phthalazin-1-one (6.4 g, 36.3 mmol), prepared as described in Example 3, in acetonitrile (65 mL), with stirring and under a dry N ₂ atmosphere at room temperature, phosphorous oxychloride (6.8 mL, 73 mmol) was added dropwise, then the mixture was heated to reflux. After 3 h, the mixture was concentrated, taken up in water, neutralized with NaHCO ₃ and extracted several times with CH ₂ Cl ₂ . The organic phases were washed with water, dewatered and concentrated to give 6.94 g of the title compound (yield: 98.3%). Melting point: 181-183 ° C.

¹ H-NMR (CDCl ₃ ) δ: 9.33 (s, 1H); 8.21-7.20 (m, 3H); 4.00 (s, 3H).

Example 5

- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazine (Compound 1)

A solution of 3,5-dichloro-4-methylpyridine (11.32 g, 70 mmol) in dry DMF (100 mL) was stirred under N ₂ at room temperature, then 60% NaH in oil (2.8 g, 70 mmol). After 1 h at room temperature, a solution of 1-chloro-6-methoxy-phthalazine (6.8 g, 35 mmol), prepared as described in Example 4, in DMF (250 mL) was added. The mixture was left at room temperature for 20 hours, then poured into ice-water (pH = 8) and extracted several times with CH 2 Cl 2. The organic phases were washed with water, dewatered and dried to give a solid which was triturated in isopropyl ether (100 mL) and filtered. The mother liquors were dried and the residue was purified by flash chromatography with ethyl acetate to give 8.9 g of the title compound (yield: 79.5%). Melting point: 173-175 ° C.

¹ H-NMR (CDCl ₃ ) δ: 9.32 (s, 1H); 8.50 (s, 2 H); 8.11 (d, 1H, J = 9.0Hz); 7.56 (dd, IH); 7.20 (d, 1H, J = 2.6Hz); 4.90 (s, 2 H); 4.00 (s, 3H).

Example 6

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxyphthalazine-3-oxide (compound 2)

To a solution of 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (1 g, 3.13 mmol), prepared as described in Example 5, in dry CH 2 Cl 2 (15 mL) was stirred and m-chloroperbenzoic acid (0.81 g, 4.7 mmol) was added to N ₂ atmosphere at room temperature. After 1 h, the mixture was washed with 10% NaOH and water, dewatered and dried. The residue was purified by flash chromatography (eluent: ethyl acetate 100%, then ethyl acetate / CH 3 OH, 8: 2) to give a solid that crystallized from acetic acid / ethyl acetate to give 0.32 g of the title compound (yield: 31%). Melting point: > 230 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.70 (s, 2H); 8.66 (s, 1 H); 8.37 (d, 1H, J = 9.1Hz); 7.42 (dd, IH); 7.34 (d, 1H, J = 2.4Hz); 4.91 (s, 2 H); 3.94 (s, 3H).

Example 7

1- (3,5-Dichloro-1-oxopyridin-4-ylmethyl) -6-methoxyphthalazine-3-oxide (compound 3)

To a solution of 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (1 g, 3.12 mmol), prepared as described in Example 5, in dry CHCl 3 (15 mL), with stirring and atmosphere N ₂ at room temperature, m-chloroperbenzoic acid (1.62 g, 9.37 mmol) was added and the mixture was refluxed. After 3 h the mixture was cooled and the solid was collected. The mother liquors were dried and the residue was purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH 3 OH 95: 5 then 9: 1) to give 0.68 g of the title compound (yield: 62%). Melting point: > 230 ° C.

¹ H-NMR (CDCl 3) δ: 8.41 (s, 1H); 8.18 (s, 21H); 8.00 (d, 1H, J = 9.2Hz); 7.32 (dd, IH); 6.93 (d, 1H, J = 2.5Hz); 4.72 (s, 2 H); 3.98 (s, 3H).

Example 8

1- 2- (4,4-Dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxyphenyl-propan-1-one

To a solution of 2- (4-methoxyphenyl) -4,4-dimethyl-4,5-dihydrooxazole (4.5 g, 21.9 mmol), prepared as described in Example 1, in dry ethyl ether (90 mL), stirring and under N ₂ at 0 ° C, 2.5 M n-butyllithium (9.6 mL, 24.1 mmol) in hexane was added. The reaction was allowed to proceed for 4 hours. In another flask, a suspension was prepared

26 ZnCl ₂ (4.48 g, 33 mmol) in dry ethyl ether (60 mL) was added a solution of the lithium derivative at 0 ° C. At the end of the addition, the mixture was left at room temperature for 1.5 hours, cooled to 0 ° C, and palladium acetate (0.25 g, 1.09 mmol), triphenylphosphine (0.57 g, 2.19 mmol) was added gradually, and after 5 min propanoyl chloride (2.13 g, 23.02 mmol). The mixture was allowed to stand for 20 hours, then poured into ice-water and extracted with ethyl acetate. The organic phase was dewatered, decolourised and dried to give 6.6 g of the title compound (yield: 75%).

¹ H-NMR (CDCl 3) δ: 7.87-6.73 (m, 3H); 4.00 (s, 2 H); 3.80 (s, 3H); 2.71 (q, 2 H); 1.31 (s, 6H); 1.16 (t, 3H, J = 7.4Hz).

Example 9

4-Ethyl-6-methoxy-2H-phthalazin-1-one

A solution of 1- [2- (4,4-dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxy-phenyl] -propan-1-one (6.4 g, 15.9 mmol), prepared according to of the description given in Example 8, in 110 mL of a mixture of water (10 mL), ethanol (100 mL) and concentrated H ₂ SO ₄ (8 mL) made up to 200 mL with ethanol, was stirred under N ₂ under reflux for 20 h, then concentrated, taken up in water and extracted several times with ethyl ether. The organic phases were washed with water, dehydrated and dried to give an oil which was dissolved in acetic acid, and hydrazine monohydrate (1.56 g, 31.12 mmol) in acetic acid (10 mL) was added at 15 ° C. After the addition was complete, the mixture was heated to 80 ° C and concentrated to a small volume after 20 hours, taken up in water, neutralized with NaHCO 3 and extracted several times with CHCl ₃ . The organic phases were drained and dried to give a residue which was purified by flash chromatography (eluent: hexane / ethyl acetate 9: 1 then 7: 3). 1.34 g of the title compound was obtained (yield: 44%). Melting point: 230-235 ° C.

··· · · · · · · · · · · · · · · · · · · · · · · · · · ·

- 27 ¹ H-NMR (CDCl₃) δ: 12.3 (s-broad, 1H); 8.18-7.27 (m, 3H); 3.92 (s, 3H); 2.93 (q, 2 H); 1.24 (t, 3H, J = 7.5Hz).

Example 10

1-Chloro-4-ethyl-6-methoxyphthalazine

To a suspension of 4-ethyl-6-methoxy-2/7-phthalazin-1-one (1.3 g, 6.76 mmol), prepared as described in Example 9, in dry acetonitrile (20 mL) was stirred and under N ₂ atmosphere at room temperature added phosphorus pentoxide (2.07 g, 13.52 mmol). The mixture was heated to reflux and after 2 hours cooled, concentrated to small volume, taken up in water, added Na ₂ CO 3 in excess and extracted three times with CH ₂ Cl ₂ . The organic phases were drained and dried to give 1.34 g of the title compound (yield: 89%).

¹ H-NMR (CDCl 3) δ: 8.20-7.54 (m, 3H); 4.01 (s, 3H); 3.30 (q, 2 H); 1.36 (t, 3H, J = 7.4Hz).

Example 11

1- (3,5-Dichloro-pyridin-4-ylmethyl) -4-ethyl-6-methoxy-phthalazine (Compound 4)

Using essentially the same procedure described in Example 5, starting with 3,5-dichloro-4-methylpyridine (1.75 g, 10.77 mmol), dry DMF (20 mL), 60% NaH in oil (0.26 g, 10.77 mmol) and 1-chloro-4-ethyl-6-methoxyphthalazine (1.2 g, 5.39 mmol), prepared as described in Example 10, in DMF (25 mL) was 0.66 g of the title compound was obtained (yield: 36%). M.p .: 136-138 ° C.

- 28 9 9 99

9999

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 8.12 (d, 1H, J = 9 Hz); 7.52 (dd, IH); 7.33 (d, 1H, J = 2.5Hz); 4.85 (s, 2 H); 4.00 (s. 3H); 3.27 (q, 2 H); 1.47 (t, 3H, J = 7.3Hz).

Example 12 [2- (4,4-Dimethyl-4,4-dihydrooxazol-2-yl-5-methoxyphenyl-phenyl) -methanone

To a solution of 2- (4-methoxyphenyl) -4,4-dimethyl-4,5-dihydrooxazole (4.5 g, 21.9 mmol), prepared as described in Example 1, in dry ethyl ether (90 mL), stirring and in N ₂ at 0 ° C, 2.5 M n-butyllithium (9.6 mL, 24.1 mmol) in hexane was added, and the reaction was allowed to proceed for 4 h. In another flask, a ZnCl ₂ slurry (4, 48 g, 33 mmol) in dry ethyl ether (60 mL) was added dropwise a solution of the lithium derivative at 0 ° C, and after 1 h benzoyl chloride (3.24 g, 23.02 mmol). The mixture was allowed to stand at room temperature for 3 hours, then bis (triphenylphosphine) palladium chloride (280 mg) was added and after 3 days the reaction mixture was poured into water / ice and extracted with ethyl acetate. The organic phases were washed with water, dewatered and dried to give 3.5 g of the title compound (yield: 52%).

¹ H-NMR (CDCl ₃ ) δ: 7.86-6.95 (m, 8H); 3.84 (s, 3H); 3.51 (s, 2 H); 0.98 (s, 6H).

Example 13

6-Methoxy-4-phenyl-2H-phthalazin-1-one

A suspension of [2- (4,4-dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxy-phenyl] -phenylmethanone (3.4 g, 11 mmol), prepared as described in Example 12, in 3N HCl (45 mL) was refluxed with stirring for 2 days, then 36% HCl (10 mL) was added. After stirring, the mixture was cooled and extracted several times with CHCl ₃ . The organic phases were drained and dried. The residue was dissolved in acetic acid and treated with hydrazine monohydrate (0.96 g, 19.24 mmol) in acetic acid (7 mL). The mixture was heated to 80 ° C in dry N ₂ and stirred for 20 hours, then concentrated to small volume, poured into water, neutralized with excess NaHCO ₃ and extracted with CHCl ₃ . The organic phases were dewatered and dried to give 0.6 g of the title compound (yield: 37%). Melting point: 230-235 ° C.

¹ H-NMR (DMSO) δ: 12.71 (s, 1H); 8.29-7.00 (m. 8H); 3.8 (s, 3 H).

Example 14

1-Chloro-6-methoxy-4-phenyl-phthalazine

Using the same procedure as described in Example 4, starting from 6-methoxy-4-phenyl-2H-phthalazin-1-one (0.6 g, 2.38 mmol), prepared as described in Example 13, acetonitrile (10 mL) and phosphorus pentoxide (0.44 mL, 4.76 mmol) gave 0.4g of the title compound (yield: 62.5%).

¹ H-NMR (DMSO) δ: 8.33-7.26 (m, 8H); 3.88 (s, 3H).

Example 15

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-4-phenylphthalazine (Compound 5)

Following the same procedure as described in Example 5, starting from 3,5-dichloro-4-methylpyridine (0.45 g, 2.81 mmol) in dry DMF (10 mL), 60% NaH (67 mg, 2, 81 mmol) and 1-chloro-6-methoxy-4-phenylphthalazine (0.38 g, 1.4 mmol), prepared as described in Example 14, in DMF (10 mL) gave 0.26 g of the title compound ( yield: 47.3%). Mp .: 206-208 ° C.

3030 ··· 9 9 ¹ H-NMR (CDCl 3) δ: 8.51 (s, 2H); 8.22-7.39 (m, 8H); 4.94 (s, 2 H); 3.87 (s, 3H).

Example 16 2- (4,4-Dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxy-phenyl-phenyl-methanol

Magnesium shavings to prepare Grignard reagent (0.12 g, 4.71 mmol) in ethyl ether (8 mL) were placed under N ₂ and the mixture was stirred at room temperature. Ethyl bromide (2 drops) was added, followed by slow addition of bromobenzene (0.52 mL, 4.93 mmol) in ethyl ether (10 mL) and 1,2-dibromoethane (2 drops). The temperature of the mixture was raised to reflux and maintained for 1 hour. 2- (4,4-Dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxybenzaldehyde (1 g, 4.29 mmol) was added to the mixture. ), prepared as described in Example 2, and the mixture was refluxed for an additional hour, then cooled and poured into ice / water and extracted several times with ethyl acetate. The organic phases were washed with water, dewatered and dried to give 1.44 g of the title compound (yield: 98%).

¹ H-NMR (CDCl 3) δ: 8.23 (s-broad, 1H); 7.80-6.67 (m, 8H); 5.86 (s-broad, 1H); 4.15- 3.79 (m, 2H); 3.76 (s, 3H); 1.31 and 0.96 (2s, 6H).

Example 17

1- [2- (4,4-Dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxyphenyl] -5-phenylpentan-1-ol

To a solution of 2- (4-methoxyphenyl) -4,4-dimethyl-4,5-dihydrooxazole (1.85 g, 9 mmol), prepared as described in Example 1, in dry ethyl ether (37 mL) at 0 ° C was added 2.5 M n-butyllithium (4 mL, 9.9 mmol) in hexane, maintaining the temperature below 5 ° C. The mixture was left at 0 ° C for 4 hours, a solution of 5-phenylpentanal (3.25 g, 20 mmol) in dry ethyl ether (10 mL) was added dropwise, keeping the temperature below 5 ° C. The mixture was kept at room temperature for 2 days, then poured into water / ice. The phases were separated and the aqueous phase was extracted again with ethyl ether. The organics were decolourised with charcoal, dewatered and concentrated to give an oil which was purified by flash chromatography (eluent: petrolatum / ethyl acetate 8: 2 then 7: 3) to give 1.17 g of the title compound (yield: 35%) .

¹ H-NMR (CDCl ₃ ) δ: 7.83-6.75 (m, 8H); 4.65-4.58 (m, IH); 4.124.02 (m, 2 H); 3.81 (s, 3H); 2.63-2.55 (m, 2 H); 2.00-1.28 (m, 6H); 1.36 (s, 6H).

Example 18

1- [2- (4,4-Dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxyphenyl] -5-phenylpentan-1-one

To a solution of 1- [2- (4,4-dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxyphenyl] -5-phenylpentan-1-ol (0.62 g, 1.7 mmol) prepared as described in Example 17 in DMSO (10 mL) was added triethylamine (35 mL) and after 30 min sulfur trioxide pyridine complex (1.62 g, 10.2 mmol). After 6 h, the mixture was diluted with 10 volumes of water and extracted twice with ethyl ether. The organic phases were washed with water, decolourised with charcoal, dewatered and concentrated. The residue was purified by flash chromatography (eluent: petrolatum / ethyl acetate 7: 3) to give 0.37 g of the title compound (yield: 60%).

¹ H-NMR (CDCl 3) δ: 7.79-6.73 (m, 8H); 3.96 (s, 2 H); 3.81 (s, 3H); 2.78-2.58 (m, 4H); 1.82-1.59 (m, 4H); 1.28 (s, 6H).

- 32 · · · · · · · · · · · · · · · · · · · · · · · · ·

Example 19

6-Methoxy-4- (4-phenylbutyl) -2,7-phthalazin-1-one

A solution of 1- [2- (4,4-dimethyl-4,5-dihydrooxazol-2-yl) -5-methoxyphenyl] -5-phenylpentan-1-one (0.37 g, 1 mmol), prepared as as described in Example 18, in 5 mL of a mixture of water (0.5 mL), ethanol (5 mL), concentrated H 2 SO 4 (0.4 mL) made up to 10 mL with ethanol, was stirred under reflux for 20 h, then concentrated small volume, taken up in water, extracted with ethyl ether and the ether phase was washed with water, dewatered and dried. The obtained oil was dissolved in acetic acid (10 mL) and a solution of hydrazine monohydrate (0.45 mL, 3 mmol) in acetic acid (2 mL) was added dropwise under N 2 atmosphere. The mixture was stirred at 80 ° C for 3 hours, then at room temperature overnight and at 80 ° C for an additional 5 hours, cooled, dried, taken up in water, basified with NaOH, and extracted several times with ethyl acetate. The organic phases were washed with water / NaCl, dewatered and concentrated. The residue was purified by flash chromatography (eluent: petroleum / ethyl acetate 6: 4) to give 0.09 g of the title compound (yield: 29%).

¹ H-NMR (CDCl 3) δ: 10.03 (s-broad, 1H); 8.40-7.06 (m, 8H); 3.91 (s, 3H); 2.93-2.63 (m, 4H); 1.88-1.71 (m, 4H).

Example 20

1-Chloro-6-methoxy-4- (4-phenylbutyl) -phthalazine

Using the same procedure as described in Example 10, starting from 6-methoxy-4- (4-phenylbutyl) -2H-phthalazin-1-one (0.2 g, 0.65 mmol), prepared as described in Example 19, in dry acetonitrile (10 mL) and POCl ₃ (0.31 g, 1.95 mmol) afforded 0.22 g of the title compound (stoichiometric yield).

¹ H-NMR (CDCl 3) δ: 8.20-7.13 (m, 8H); 3.93 (s, 3H); 3.29-3.22 (m, 2 H); 2.72-2.64 (m, 2 H); 1.98-1.72 (m, 4H).

- 33 · · 33 33 33 33 33 33

Example 21

1- (3 _l 5-dichloropyridine-4-ylmethyl) -6-methoxy-4- (4-phenylbutyl) phthalazine (Compound 6)

Using the same procedure as described in Example 5, starting from 3,5-dichloro-4-methylpyridine (0.23 g, 1.4 mmol) in dry DMF (10 mL), 55% NaH in oil (0.061 g, 1.4 mmol) and 1-chloro-6-methoxy-4- (4-phenyl-butyl) -phthalazine (0.23 g, 0.7 mmol), prepared as described in Example 20, in DMF (5 mL), 0.082 g of the title compound was obtained as an amorphous hygroscopic solid (yield: 21%).

¹ H-NMR (CDCl 3) δ: 8.79-7.94 (m, 3H); 8.72 (s, 2 H); 7.27-7.10 (m, 5H); 5.70 (broad, H ⁺ ); 5.12 (s. 2H); 4.09 (s, 3H); 3.48 (t-broad); 2.60 (t-broad, 2H); 1.88-1.61 (m, 4H).

Example 22

4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (compound 7)

A suspension of 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (23.5 g, 73.4 mmol) prepared as described in Example 5 and PtC > 2 hydrate (0.5 g, 2 (2 mmol) in THF (600 mL) was charged to a hydrogenation reactor at room temperature and 20 psi pressure. After 22 h, the mixture was filtered through celite and dried to give 23.8 g of the title compound (stoichiometric yield). Melting point: 181-183 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.45 (s, 2H); 7.37-6.63 (m. 3H); 4.20 (s, 2 H); 4.08 (s, 2 H); 3.83 (s, 3H).

- 34 · · 34 34 34 34 34 34 34 34 34

Example 23

144- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1H-phthalazin-2-yl-ethanone (Compound 8)

To a solution of 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazine (1 g, 3.12 mmol), prepared as described in Example 5, in glacial acetic acid (30 mL), with stirring In N ₂ , 10% Pd / C was added in catalytic amount, and the solution was charged to a hydrogenation reactor at 0.4 MPa. After 2 days, the mixture was filtered and dried. The residue was purified by flash chromatography (eluent: hexane / ethyl acetate 3: 7) to give 0.5 g of the title compound (yield: 50%). Melting point: 186-188 ° C.

ET As an alternative to the previous synthesis, the title compound was prepared from a solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (2 g, 6.2 mmol), prepared as described above. as described in Example 22, in dry THF (60 mL) as starting material, with stirring in dry N ₂ at 0 ° C. To the mixture was added triethylamine (1.57 g, 15.5 mmol) then acetyl chloride (0.44 mL, 6.2 mmol). After 10 min, the mixture was brought to room temperature and maintained at this temperature for 2.5 hours, then poured into water / ice and extracted several times with CH ₂ Cl ₂ . The organic phases were washed with 5% NaOH and water, dewatered and dried to give a solid that crystallized from acetonitrile (70 mL) to give 1.95 g of the title compound having the same physicochemical properties as the compound obtained in para. AND.

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 7.46 (d, 1H, J = 8.6Hz); 6.88 (dd, IH); 6.70 (d, 1H, J = 2.4Hz); 4.85 (s, 2 H); 4.29 (s, 2 H); 3.84 (s, 3H); 1.84 (s, 3H).

- 35 ·· 94 • 4 9 4 • 9 · 9 · 9

4 4

9494

4 • 9 4

9 4 4 • 4 9444

4 4 *

9 4

4

4 9

4

4

Example 24

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1H-phthalazine-2-carboxylic acid amide (Compound 9)

To a solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (0.4 g, 1.24 mmol), prepared as described in Example 22, in glacial acetic acid ( 8 mL), with stirring and under an atmosphere of dry N ₂ at room temperature, potassium cyanate (0.2 g, 2.48 mmol) was added and the reaction mixture was allowed to stand for 20 hours, then poured into 10% NaOH in excess and extracted several times with CH ₂ Cl ₂ . The organic phases were washed with water, dewatered and concentrated to give a residue which was purified by flash chromatography (eluent: hexane / ethyl acetate 3: 7) to give 0.36 g of the title compound (yield: 80%). Melting point: > 230 ° C.

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 7.47-6.69 (m. 3H); 5.01 (broad, 2H); 4.82 (s, 2 H); 4.29 (s, 2 H); 3.85 (s, 3H).

Example 25

1- [4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1 H -phthalazin-2-yl] -propan-1-one (Compound 10)

To a solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (1 g, 3.1 mmol), prepared as described in Example 22, in THF (35 mL) ), with stirring under a dry N ₂ atmosphere at room temperature, triethylamine (1.08 mL, 7.76 mmol) was added dropwise, followed by propionyl chloride (0.323 mL, 3.72 mmol) at 0 ° C. After 10 min, the mixture was brought to room temperature and after 3 h it was poured into water / ice and THF was evaporated. The mixture was extracted several times with CH ₂ Cl ₂ and the organic phases were washed with 5% NaOH and water, dewatered and dried to give a solid that crystallized from acetonitrile (50 mL). 1 g of the title compound was obtained (yield: 86%). M.p .: 195-197 ° C.

36 ¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 7.48-6.70 (m, 3H); 4.85 (s, 2 H); 4.29 (s, 2 H); 3.84 (s, 3H); 2.15 (q, 2 H); 0.87 (t, 3H, J = 7.5Hz).

Example 26

1-í4- (3 _l 5-dichlorpvridin-4-ylmethyl) -7-methoxy-1/7-phthalazin-2-vl1-2-methylpropan-1-one (Compound 11)

Using essentially the same procedure described in Example 25, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (1 g, 3.1 mmol) prepared as described above. Example 22, in dry THF (35 mL), triethylamine (1.08 mL, 7.76 mmol) and isobutanoyl chloride (0.39 mL, 3.72 mmol) gave 1 g of the title compound (yield: 83%) . Melting point: 210-212 ° C.

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 7.46 (d, 1H, J = 8.6Hz); 6.88 (dd, IH); 6.71 (d, 1H, J = 2.6Hz); 4.83 (s, 2 H); 4.29 (s, 2 H); 3.84 (s, 3H); 2.80-2.60 (m, IH); 0.83 (d, 6H, J = 6.8Hz).

Example 27 [4- (3,5-Dichloro-pyridin-4-ylmethoxy-7-methoxy-1 H -phthalazin-2-yl) -phenyl-methanone (Compound 12)

Using essentially the same procedure described in Example 25, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (1 g, 3.1 mmol) prepared as described above. in Example 22, in dry THF (35 mL), triethylamine (1.08 mL,

7.76 mmol) and benzoyl chloride (0.43 mL, 3.72 mmol) gave 1.1 g of the title compound (yield: 85%). Melting point: 190-192 ° C.

¹ H-NMR (CDCl 3) δ: 8.24 (s, 1H); 7.50-6.77 (m, 8H); 5.03 (s, 2 H); 4.22 (s, 2 H); 3.87 (s, 3H).

- 37 ··

Example 28

1- [4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1 H -phthalazin-2-yl] -phenyl-ethanone (Compound 13)

Using essentially the same procedure as described in Example 25, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (1 g, 3.1 mmol), prepared as was described in Example 22, in dry THF (35 mL), triethylamine (1.08 mL,

7.76 mmol) and phenylacetyl chloride (0.49 mL, 3.72 mmol) gave 0.9 g of the title compound (yield: 66.2%). M.p .: 143-145 ° C.

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 7.47 (d, 1H, J = 8.6Hz); 7.2-6.85 (m, 6H) + 6.71 (d, 1H, J = 2.6 Hz); 4.88 (s, 2 H); 4.41 (s, 2 H); 3.84 (s, 3H); 3.58 (s, 3H).

Example 29

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1H-phthalazine-2-carbaldehyde (Compound 14)

To a solution of formic acid (0.123 mL, 3.255 mmol) in dry THF (40 mL), with stirring and under a dry N ₂ atmosphere at room temperature, was added 1,1'-carbonyldiimidazole (0.53 g, 3.255 mmol) and the mixture 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (1 g, 3.1 mmol), prepared as described in Example 22, was added portionwise. After 6.5 hours, another portion of the formic acid / carbonyldiimidazole mixture (approximately 1/3 of the previous dose) was added and the mixture was stirred at room temperature for 1 hour and allowed to stand overnight. The mixture was poured into water / ice, the THF was evaporated and the solution was extracted several times with CH ₂ Cl ₂ . The organic phase was washed with KHSO ₄ , then with NaHCO ₃ and water, dewatered and concentrated. The residue was crystallized from acetonitrile

-38 0 0 00 0 0 0

0 0 • 000 • 00

0000 • 0 0 • 0 0 0 • 0

Φ

0 0 0

0 00 0 (40 mL) to give 1 g of the title compound (yield: 92.6%). Melting point: 165 - 167 ° C ¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 8.29 (s, 1 H); 7.47 (s, 1H, J = 8.6Hz); 6.89 (dd, IH); 6.71 (d, 1H, J = 2.6Hz); 4.82 (s, 2 H); 4.29 (s, 2 H); 3.85 (s, 3H).

Example 30

4- (3,5-Dichloro-pyridin-4-ylmethyl) -2-methanesulfonyl-7-methoxy-1,2-dihydrophthalazine (Compound 15)

Using essentially the same procedure as described in Example 25, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (1 g, 3.1 mmol) prepared as described above. in Example 22, in dry THF (30 mL), triethylamine (1.08 mL,

7.76 mmol) and methanesulfonyl chloride (0.29 mL, 3.72 mmol) yielded 0.9 g of the title compound (yield: 75.6%). Melting point: 171-173 ° C.

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 7.48-6.73 (m, 3H); 4.45 and 4.31 (2s, 4H); 3.85 (s, 3H); 2.75 (s, 3H).

Example 31

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1H-phthalazine-2-carboxylic acid methyl ester (Compound 16)

A solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydro-phthalazine (1 g, 3.1 mmol), prepared as described in Example 22, in THF (35 mL) , under stirring in a dry N ₂ atmosphere at room temperature, was mixed with 1,1'-carbonyldiimidazole (0.55 mL, 3.41 mmol). The mixture was refluxed for 1 h. Dry CH ₃ OH (2 mL) was added and heating continued for an additional 2.5 h. The mixture was cooled, poured into water / ice and extracted with CH ₂ Cl ₂ ·

- 39 ·· 99 • 9 9 • 9 9 • 99 • 9 9999

9 9

9

9999 • 99 • 9

9

9 9

9 ·· ·

The organic phases were washed with KHSO4, then with 5% NaOH, and finally with water, dewatered and concentrated to give a solid which was taken up in CH3OH with a catalytic amount of NaOCH3. The solution was refluxed for 1.5 h, cooled, CH ₃ OH was evaporated, the residue was taken up in water and CH 2 Cl 2. The organic phases were washed with NaOH and water, dewatered and concentrated to give a solid which was triturated with CH ₃ OH. 0.6 g of the title compound was obtained (yield: 52%). Melting point: 184-186 ° C.

¹ H-NMR (CDCl 3) δ: 8.46 (s, 2H); 7.36 (d, 1H, J = 8.6Hz); 6.83 (dd, IH); 6.69 (d, 1H, J = 2.6Hz); 4.77 (s, 2 H); 4.32 (s, 2 H); 3.82 (s, 3H); 3.70 (s, 3H).

Example 32

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1H-phthalazine-2-carboxylic acid methylamide (Compound 17)

Using essentially the same procedure as described in Example 31, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydrophthalazine (1 g, 3.1 mmol) prepared as described above. in Example 22, in dry THF (35 mL), 1,1'-carbonyldiimidazole (0.55 g, 3.41 mmol) and 8.03 M methylamine in ethanol (2 mL) gave 1 g of the title compound ( yield; 86%). Melting point: 203 - 205 ° C.

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 7.41 (d, 1H, J = 8.6Hz); 6.86 (dd, IH); 6.7 (d, 1H, J = 2.6Hz); 5.63-5.55 (m, IH); 4.80 (s, 2 H); 4.27 (s, 2 H); 3.84 (s, 3H); 2.69 (d, 3H, J = 5Hz).

-40 • 4 · 4 · 4 · 4444 49 • 4

9 4 * 443 »

Example 33

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1H-phthalazine-2-carboxylic acid dimethylamide (Compound 18)

A solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1,2-dihydro-phthalazine (1 g, 3.1 mmol), prepared as described in Example 22, in dry THF (35 mL) ), under stirring in a dry N ₂ atmosphere at room temperature, was mixed with 1, T-carbonyldiimidazole (0.6 g, 3.72 mmol), then the mixture was refluxed for 3 hours, cooled, 5.6 was added M dimethylamine in ethanol (5 mL) and the mixture was again refluxed. After 44 h, the mixture was cooled, poured into water / ice and extracted several times with CH 2 Cl 2. The organic phases were washed with 5% NaOH, then with water, dewatered and concentrated. The residue was purified by flash chromatography (eluent: hexane / ethyl acetate 1: 1), then crystallized from acetonitrile (30 mL) to give 0.44 g of the title compound (yield: 36.4%). Melting point: 160-162 ° C.

¹ H-NMR (CDCl 3) δ: 8.45 (s, 2H); 7.43 (d, 1H, J = 8.5Hz); 6.88 (dd, IH); 6.71 (d, 1H, J = 2.6Hz); 4.49 (s, 2 H); 4.27 (s, 2 H); 3.84 (s, 3H); 2.64 (s, 6H).

Example 34

6-Methoxy-3H-isobenzofuran-1-one

Concentrated HCl (1 L) was mixed with 40% w / v formaldehyde (65 mL, 0.86 mol) with vigorous stirring, then 3-methoxybenzoic acid (100 g, 0.66 mol) was added and the mixture was heated to 100 ° C while controlling gas formation for 30 min. Cooling of the mixture gave a precipitate which was filtered and stored while the mixture was washed with water and then with 5% NaOH. The new precipitate was extracted twice with CH 2 Cl 2, the extract was dewatered, concentrated, combined with a previously filtered solid and both; · · · · · ·

2 · · · ·. ·······

The portions were dissolved in CH 2 Cl 2 and treated with diethylamine (120 mL, 1.15 mol). After 24 h the mixture was extracted with 10% HCl and the phases were separated and extracted with CH 2 Cl 2. The organic phase was washed with 10% NaOH, decolourised with charcoal, dewatered and concentrated. The residue was dissolved in CH 2 Cl 2 and treated with 10% HCl for 30 min with stirring. The organic phase was washed with water, dewatered and dried. The residue was dissolved in CH ₂ Cl ₂ and treated with 10% NaOH with stirring for 30 min. The organic phase was washed with water, dewatered and concentrated to give a solid which was crystallized from CH ₃ OH / H ₂ O 65:35 and dried at 50 ° C over P2O 5, then crystallized again from CH ₃ OH / H ₂ O 6: 4. 35.28 g of the title compound is obtained (yield: 32%). Melting point: 115 - 117 ° C ¹ H-NMR (CDCl 3) δ: 7.37-7.20 (m, 3H); 5.21 (s); 3.85 (s, 3H).

Example 35

3-Bromo-6-methoxy-3 H -isobenzofuran-1-one

6-Methoxy-3H-isobenzofuran-1-one (35.28 g, 0.215 mol), prepared as described in Example 34 and suspended in CCl ₄ (350 mL) under N ₂ was mixed with N-bromosuccinimide (40 g) (0.225 mol) and benzyl peroxide in a catalytic amount and the mixture was slowly brought to reflux. After 2.5 hours, heating was stopped and the mixture was allowed to stand overnight at room temperature. Additional catalyst was added and the mixture was heated for an additional 3.5 h. The mixture was cooled in ice, filtered through celite, washed well with CCl 4 and dried to give 41 g of the title compound (yield: 78%).

¹ H-NMR (CDCl 3) δ: 7.50-7.25 (m, 4H); 3.87 (s, 3H).

9

- 42 9 9 • 9 · «····

9

9 9

9

Example 36 (5-Methoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenylphosphonium bromide

3-Bromo-6-methoxy-3/7-isobenzofuran-1-one (41 g, 0.169 mol), prepared as described in Example 35, suspended in dry acetonitrile (205 mL) under N 2 was mixed with triphenylphosphine (42 g). g, 0.16 mol). The mixture was refluxed and after about 3 hours it was cooled and concentrated to give a solid which was mixed with ethyl ether, filtered and dried in vacuo. 74 g of the title compound are thus obtained (yield: 84%).

¹ H-NMR (CDCl ₃ ) δ: 9.63 (s, 1H); 7.84-7.75 (m, 15H); 7.09-6.91 (m, 3H); 3.77 (s, 3H).

Example 37

3- (3,5-Dichloropyridin-4-ylmethylene) -6-methoxy-3H-isobenzofuran-1-one

To a suspension of N2 (5-methoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenylphosphonium bromide (74 g, 0.134 mol), prepared as described in Example 36 and 3,5-dichloropyridine-4-carbaldehyde (23.6 g, 0.134 mol) in CH 2 Cl 2 (500 mL) was added dropwise triethylamine (18.5 mL, 0.134 mol), adjusting the temperature using a water bath. The mixture was stirred overnight, then cooled and treated with 5% HCl. The phases were separated and the acidic phase was extracted with CH 2 Cl 2, washed with H ₂ O / NaCl, decolourised with charcoal, dewatered and concentrated under high vacuum. 85.4 g of crude material was obtained, which was used as such in the next step. A sample of the crude material was purified by flash chromatography (eluent: hexane / ethyl acetate 1: 1).

¹ H-NMR (CDCl 3) δ: 8.60 (s, 2H); 7.77-6.68 (m, 4H); 3.80 (s, 3H).

-43 · · 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43 43

Example 38

4- (3,5-dichloro-pyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (compound 19)

To a suspension of 3- (3,5-dichloropyridin-4-ylmethylene) -6-methoxy-3/7-isobenzofuran-1-one (84.4 g, 0.126 mol), prepared as described in Example 37, in CH 3 OH (200 mL) ) under N ₂ was added hydrazine monohydrate (18.4 mL, 0.378 mol). The mixture was refluxed for 1 hour, left over the nose at room temperature and cooled with ice. The solid phase was filtered off, washed with cold CH 3 OH and dried in an oven at 50 ° C under vacuum; 33.3 g of the title compound are thus obtained (yield: 80%). Melting point: 259-262 ° C.

¹ H-NMR (CDCl 3) δ: 12.34 (s, 1H); 8.64 (s, 2 H); 8.19-7.54 (m, 3H); 4.58 (s, 2 H); 3.95 (s, 3H).

Example 39

4- (3,5-Dichloropyridin-4-ylmethyl) -2-ethyl-7-methoxy-2 H -phthalazin-1-one (compound 20)

A suspension of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (0.52 g, 1.55 mmol), prepared as described in Example 38, in DMF (10 mL) was treated with NaH (0.057 g, 1.55 mmol) and after 1 h with ethyl iodide (0.125 mL, 1.55 mmol). After one night, the mixture was diluted with water (10 volumes) and extracted twice with ethyl acetate. The organic phase was dewatered and concentrated to give a solid which was purified by flash chromatography (eluent: ethyl acetate / petrolatum 3: 7). There was thus obtained 0.44 g of the title compound (yield: 78%). Melting point: 150.2 151.2 ° C.

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 7.86-7.36 (m, 3H); 4.51 (s, 2 H); 4.01 (q, 2H, J = 7.02 Hz); 3.96 (s, 3H); 1.13 (t, 3 H).

-44 9 9 9

9

9

9

9999

9

9 9

9 9 9 • 9999 9

9 9

9

Example 40

2-Benzyl-4- (3,5-dichloro-pyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (Compound 21)

Using essentially the same procedure as described in Example 39, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (1 g, 2.55). mmol), prepared as described in Example 38, in DMF (10 mL), NaH (0.1 g, 2.55 mmol) and benzyl chloride (0.32 mL, 2.8 mmol) yielded 0.6 g in of the title compound (yield: 55%). Melting point: 134-135 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.50 (s, 2H); 7.85-7.35 (m, 3H); 7.22-7.12 (m, 5H); 5.10 (s. 2H); 4.50 (s, 2 H); 3.95 (s, 3H).

Example 41

4- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxy-2-phenethyl-2H-phthalazin-1-one (compound 22)

Using essentially the same procedure as described in Example 39, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (1 g, 2.55 mmol), prepared as as described in Example 38, in DMF (10 mL), NaH (0.1 g, 2.55 mmol) and (2-chloroethyl) -benzene (0.36 mL, 2.8 mmol) yielded 0.7 g of the title compound (yield: 62%). Melting point: 147.5-148.5 ° C.

¹ H-NMR (CDCl 3) δ: 8.52 (s, 2H); 7.84-7.37 (m, 3H); 7.21-7.06 (m, 5H); 4.51 (s, 2 H); 4.26 - 4.19 (m, 2H); 3.96 (s, 3H); 2.92-2.84 (m, 2 H).

··

-45 9 9 9 9 9 9 9 9 9 9 9 9 · ·

Example 42

4- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxy-2- (5-phenylpentyl) -2H-phthalazin-1-one (compound 23)

Using essentially the same procedure as described in Example 39, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (1.34 g, 3.4 mmol) ), prepared as described in Example 38, in DMF (25 mL), NaH (0.13 g, 3.4 mmol) and (5-phenylpent-1-yl) -4-methylphenylsulfonate (1.34 mL, 4). , 2 mmol), 1.1 g of the title compound was obtained (yield: 24%). Melting point: 107.9 109.9 ° C.

¹ H-NMR (CDCl 3) δ: 8.49 (s, 2H); 7.86-7.37 (m, 3H); 7.27-7.07 (mm, 5H); 4.51 (s, 2 H); 4.01-3.94 (m, 2 H); 3.97 (s, 3H); 2.55-2.17 (m, 2 H); 1.68-1.12 (m, 6H)

Example 43

4- (3,5-Dichloropyridin-4-ylmethyl) -2-methanesulfonyl-7-methoxy-2H-phthalazin-1-one (compound 24)

Using essentially the same procedure as described in Example 39, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (0.5 g, 1.27) mmol), prepared as described in Example 38, in DMF (10 mL), NaH (0.05 g, 1.27 mmol) and mesyl chloride (0.1 mL, 1.27 mmol), 0.18 g was obtained in of the title compound (yield: 35%). Melting point: 192.6-193.6 ° C.

¹ H-NMR (CDCl 3) δ: 8.52 (s, 2H); 7.86 (d, 1H, J = 2.5Hz); 7.845 (d, 1H, J = 8.7Hz); 7.48 (dd, IH); 4.55 (s, 2 H); 3.98 (s, 3H); 3.32 (s, 3H).

• ·

Example 44

[4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1-oxo-1 H -phthalazin-2-yl-1-acetic acid ethyl ester (Compound 25)

Using essentially the same procedure as described in Example 39, starting from 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (2 g, 5.1 mmol), prepared as as described in Example 38, in DMF (20 mL), NaH (0.2 g, 5.1 mmol) and ethyl bromoacetate (0.62 mL, 5.61 mmol), a crude was obtained which crystallized from the mixture isopropyl ether / acetonitrile 85: 15 (40 mL), yielding 1.12 g of the title compound (yield: 52%). M.p .: 141-142 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.48 (s, 2H); 7.85-7.39 (m, 3H); 4.688 (s, 2 H); 4.52 (s, 2 H); 4.14 (q, 2 H); 3.96 (s, 3H); 1.20 (t, 3H, J = 7.1Hz).

Example 45

4-Chloro-1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazine

To a suspension of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2H-phthalazin-1-one (10 g, 25.5 mmol), prepared as described in Example 38, in acetonitrile (300 mL) POCl 3 (22.2 mL, 230 mmol) was added and the mixture was heated to reflux. After 3 h, the solution was concentrated, taken up in CH 2 Cl 2 in water and the pH was adjusted to 7-8 with Na ₂ CO ₃ . The organic phases were decolorized with activated carbon, dehydrated and concentrated to give 10 g of the title compound (stoichiometric yield). Melting point: 156-166 ° C.

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 8.13-754 (m, 3H); 4.88 (s, 2 H); 4.04 (s, 3H).

· · · · · Λ · λ · λ λ λ ~ λ λ λ λ λ λ λ λ λ λ 7 λ · ··· ···

Example 46

4-Bromo-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine

Κ a suspension of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-2- [7-phthalazin-1-one (10 g, 25.5 mmol), prepared as described in Example 38, in acetonitrile ( 300 mL), POBr ₃ (22 g, 76.5 mmol) was added and the mixture was refluxed. After one night at reflux and the next night standing, the solution was concentrated, taken up in water, cooled in ice and basified with concentrated NaOH. The mixture was extracted three times with CH 2 Cl 2 and the organic phases were decolorized with activated carbon, dehydrated and concentrated. The crude was purified by flash chromatography (eluent: petrolatum / ethyl acetate 8: 2 then 6: 4) to give 8.4 g of the title compound (yield: 82%).

¹ H-NMR (CDCl 3) δ: 8.49 (s, 2H); 8.12-7.50 (m, 3H); 4.83 (s, 2 H); 4.04 (s, 3H).

Example 47

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-4-phenylphthalazine (Compound 5)

To ZnCl 2 (2.04 g, 15 mmol) under N ₂ in THF (100 mL) was added phenyllithium (7.5 mL, 15 mmol; 2M in cyclohexane / ethyl ether 7: 3) at 0-4 ° C. The mixture was stirred at room temperature for 1 hour. 4-Bromo-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (3 g, 7.5 mmol), prepared as described in Example 46, was added, palladium acetate (0.034 g, 0.15 mmol) and triphenylphosphine (0.08 g, 0.3 mmol) and the mixture was refluxed for 24 h, cooled and extracted with saturated NH 4 Cl solution. The organic phases were extracted again with ethyl acetate, decolourised with charcoal, dewatered and concentrated to give a solid which was purified by flash chromatography (eluent:

• · • ·

AQ · · · · · ···

- * ι · lacetát - ·········· Petrolatum / ethyl acetate 6: 4). The obtained oil, which was mixed with ethyl ether, crystallized from acetonitrile (80 mL) and was dried overnight to give 1.44 g of the title compound (yield: 55%).

The analytical data obtained for this compound were consistent with the data for the same compound obtained as described in Example 15.

Example 48

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4-thiazol-2-yl-phthalazine (Compound 26)

To a suspension of zinc (0.18 g, 2.76 mmol) in THF / toluene 2: 1 (15 mL) under stirring under N ₂ at reflux was added

2-bromothiazole (0.23 mL, 2.5 mmol). After 2 1/2 hours, 4-bromo-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (0.5 g, 1.25 mmol), prepared as described in Example 46, acetate was added. palladium (14 mg, 0.0625 mmol) and triphenylphosphine (32 mg, 0.125 mmol) to the cold mixture. The mixture was refluxed for an additional 30 min. After 21 hours, equal amounts of palladium acetate and triphenylphosphine were added. After 3 hours under reflux, the mixture was cooled and poured into a mixture of water / ice and NH ₄ Cl, then extracted first with ethyl acetate, then CH ₂ Cl ₂ . The organic phases were washed with water, dewatered and concentrated to give a solid which was purified by flash chromatography (eluent: hexane / ethyl acetate 7: 8) to give 0.26 g of the title compound (yield: 51%). Melting point: 225-230 ° C.

¹ H-NMR (CDCl 3) δ: 9.34 (d, 1H, J = 2.6 Hz); 8.51 (s, 2 H); 8,197.56 (m, 3H); 7.49 (d, IH); 4.93 (s, 2 H); 4.07 (s, 3H).

• φ φφφ φφφφφ

Example 49

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4-phenyl-ethynyl-phthalazine (Compound 27)

To a solution of 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazine (0.52 g, 1.5 mmol), prepared as described in Example 45, in dry DMF (10 mL) under N ₂ atmosphere, piperidine (1 mL, 10 mmol), phenylacetylene (0.15 g, 1.5 mmol), PdCl ₂ (0.013 g, 0.075 mmol), triphenylphosphine (0.039 g, 0.15 mmol) were added. ) and CuI (0.014 g, 0.075 mmol). The mixture was stirred for 20 h, poured into saturated NH ₄ Cl (10 volumes) and extracted twice with CH ₂ Cl ₂ , then washed with 5% HCl, dewatered and concentrated to give a solid which was purified by flash chromatography (eluent: CH ₂ Cl _{2). 2} / ethyl ether 9: 1). The solid obtained was triturated in ethyl ether to give 0.41 g of the title compound (yield: 52%). Melting point: 213.4 - 214.4 ° C (dec.).

¹ H-NMR (CDCl 3) δ: 8.51 (s, 2H); 8.15-7.37 (m, 8H); 4.91 (s, 2 H); 4.05 (s, 3H).

Example 50

1- [4- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazin-1-yl] pyrrolidin-2-one (compound 28)

To a solution of 2-pyrrolidinone (0.14 mL, 1.83 mmol) in dry DMF (10 mL) under N ₂ was added NaH (0.068 g, 1.7 mmol; 60% in oil) and the mixture was heated to 40 ° C. ° C for 1 hour, cooled and added

4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (0.5 g, 1.41 mmol), prepared as described in Example 45. The mixture was heated at 80 ° C overnight It was then poured into water (10 volumes) and extracted three times with ethyl acetate. The organic phases were decolourised with charcoal, dewatered and concentrated to give a solid which was purified by flash chromatography (eluent: ethyl acetate / petrolatum 9: 1). The solid obtained was mixed with ethyl ether to give 0.306 g of the title compound (yield: 53%). Melting point: 223-225 ° C.

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 8.14-7.23 (m, 3H); 4.87 (s, 2 H); 4.22 (t, 2H, J = 6.9Hz); 3.97 (s, 3H); 2.69 (t, 2H, J = 7.9Hz); 2.37-2.22 (m, 2 H).

Example 51

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-4-F1,2,4-triazol-1-yl-phthalazine (Compound 29)

Using essentially the same procedure described in Example 50, starting from 1,2,4-triazole (0.19 g, 2.8 mmol) in dry DMF (10 mL), NaH (0.084 g, 2.1 mmol) 60% in oil) and 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (0.5 g, 1.4 mmol), prepared as described in Example 45, was obtained 0.42 g of the title compound (yield: 77%). Melting point: 207 - 208 ° C.

¹ H-NMR (CDCl 3) δ: 9.21 (s, 1H); 8.53 (s, 2 H); 8.46 (d, 1H, J = 2.6Hz); 8.25 (s, 1 H); 8.22 (d, 1H, J = 9.1Hz); 7.66 (dd, IH); 4.94 (s, 2 H); 4.02 (s, 3H).

Example 52

3-Benzyloxy-4-methoxybenzaldehyde

A mixture of 3-hydroxy-4-methoxybenzaldehyde (100 g, 0.657 mol), ethanol (300 mL), K ₂ CO ₃ (108.9 g, 0.788 mol), benzyl chloride (86.8 mL, 0.755 mol) and NaI (5 mL). g) under an N ₂ atmosphere, was stirred under reflux for 2.5 hours, then cooled, and water (900 ml) was added. The precipitate was filtered, washed with water and then with petrolatum, then dried in vacuo at 40 ° C to give 159 g. 4 4 9 4 4 4 4445 4 4 4 ·

C 44 44 44 94 44 44 44 of the title compound which was used as such in the next step.

Example 53

3-Benzyloxy-4-methoxybenzoic acid

To a solution of KMnCl 4 (24.81 g, 0.157 mol) in water (100 mL) was added a solution of tetrabutylammonium bromide (50.61 g, 0.157 mol) in water (200 mL) with stirring, and the mixture was diluted with additional water (200 mL). ml). The solid was collected by filtration, squeezed and dissolved in pyridine (300 mL). The solution was added dropwise to a solution of 3-benzyloxy-4-methoxybenzaldehyde (38.2 g, 0.157 mol), prepared as described in Example 52, in pyridine (150 mL) in a water bath. After 3 h, the mixture was acidified to pH 1N HCl, the solid was filtered, and the mother liquors were extracted several times with CH 2 Cl 2. The organic phase was dewatered, concentrated and the residue was taken up in 1N NaOH and washed with ethyl ether. The aqueous solution was acidified and extracted twice with CH 2 Cl 2, dried, bleached with TONSIL® and concentrated to a small volume. The precipitate was filtered off to give 35.869 g of the title compound (yield: 88%).

¹ H-NMR (CDCl 3) δ: 7.78-6.69 (m, 8H); 5.18 (s, 2 H); 3.39 (s, 3H).

Example 54

3-Benzyloxy-4-methoxybenzyl chloride

A solution of 3-benzyloxy-4-methoxybenzoic acid (35.86 g, 0.139 mol), prepared as described in Example 53, in thionyl chloride (150 mL), was refluxed for 2 hours under N 2, evaporated to dryness and evaporated twice taken up in toluene (100 mL) to give 35.01 g of the title compound (yield: 98%).

¹ H-NMR (CDCl 3) δ: 7.84-6.90 (m, 8H); 5.16 (s, 2 H); 3.95 (s, 3H).

Example 55

3-Benzyloxy-N, N -diethyl-4-methoxybenzamide

To a solution of 3-benzyloxy-4-methoxybenzoyl chloride (35.01 g, 0.127 mol), prepared as described in Example 54, in CH ₂ Cl ₂ (200 mL), diethylamine (131 mL) was added dropwise at 5-10 ° C. , 92.54 g, 1.27 mol) in CH ₂ Cl ₂ (130 mL). The mixture was evaporated to dryness, dissolved in ethyl acetate, washed with water, 2% KHSO 4, again with water and NaHCO 3, dehydrated over Na ₂ SO 4 and dried. The residue was treated with petrolatum (250 mL) to give 37.65 g of the title compound (yield: 95%).

¹ H-NMR (CDCl 3) δ: 7.43-6.84 (m, 8H); 5.14 (s, 2 H); 3.88 (s, 3H); 3.31 (m, 4H); 1.07 (m, 6H).

Example 56

3-Benzyloxy-N, N -diethyl-2-formyl-4-methoxybenzamide

To a solution of 3-benzyloxy-N, N-diethyl-4-methoxybenzamide (39.54 g, 126.2 mmol) prepared as described in Example 55 and tetramethylethylenediamine (16.13 g, 138.8 mmol) ) in THF (4.250 mL), cooled to -78 ° C under N ₂ , was added dropwise 1.21M sec-butyllithium (115.64 mL, 138.8 mmol). After 2 h, DMF (43 mL, 555 mmol) was added and the mixture was left at -78 ° C for 4 h, then allowed to rise overnight. The mixture was washed with 0.4 M phosphate buffer pH 7, the organic phase was separated and the aqueous phase was extracted with ethyl ether. The organic phase was drained and dried to give a solid which was filtered off. The mother liquors were dried and the raw material was purified by flash chromatography (eluent:

9

CO ··· ··· ··· ~ OO - * · ···· ·· · ·· ·· <

ethyl acetate / petrolatum 1: 1) to give 13.76 g of the title compound (yield: 32%).

¹ H-NMR (CDCl 3 δ: 10.22 (s, 1H); 7.36-7.29 (m, 5H); 7.13 and 6.94 (2s, 2H, J = 8.3 Hz); 5.17 (s, 2H), 3.94 (s, 3H), 3.52 and 2.93 (2q, 4H), 1.26 and 0.93 (2t, H).

Example 57

3,4-dihydroxy-5-methoxy-3H-isobenzofuran-1-one

A solution of 3-benzyloxy- N, N -diethyl-2-formyl-4-methoxybenzamide (11.6 g, 34 mmol), prepared as described in Example 56, in 10% HCl (80 mL) and acetic acid (80 mL) was refluxed for 18 h. The solvents were evaporated and the crude was taken up in toluene. The solid was washed with ethyl ether and dried to give 6.66 g of the title compound (quantitative yield) which was used as such in the next step.

¹ H-NMR (CDCl 3) δ: 9.64 and 7.82 (2s broad, 2H); 7.28-7.16 (m, 2 H); 3.89 (s, 3H); 6.53 (s broad, 1H).

Example 58

5-Hydroxy-6-methoxy-2H-phthalazin-1-one

To a solution of 3,4-dihydroxy-5-methoxy-3H-isobenzofuran-1-one (6.63 g, 34 mmol), prepared as described in Example 57, in ethanol (60 mL) was added 98% hydrazine ( 8.5 ml). The mixture was heated until clear and after 5 min the formed precipitate was filtered off and mixed with 1N HCl. The mother liquors were concentrated several times and the obtained solid fractions were combined to give 4.65 g of the title compound (yield: 72%).

¹ H-NMR (CDCl 3) δ: 12.33 (m, 1H); 10.06 (s, 1 H); 8.36 (s, 1 H); 7.68 and 7.49 (2d, 2H, J = 8.6 Hz); 3.94 (s, 3H).

• ·

- 54 • 9 9 9 9 9 9 ⁹ 99 99 99 99 99

Example 59

Trifluoromethanesulfonic acid 6-methoxy-1-oxo-1,2-dihydrophthalazin-5-yl ester

To a suspension of 5-hydroxy-6-methoxy-2H-phthalazin-1-one (4.9 g, 25.5 mmol), prepared as described in Example 58, in CH 2 Cl 2 (100 mL) and pyridine (10 mL). Trifluoromethanesulfonic anhydride (4.72 mL, 28.05 mmol) was added dropwise at -5 to 0 ° C under N ₂ atmosphere. After 30 min at -5 ° C additional trifluoromethanesulfonic anhydride (0.94 mL) was added and after 30 min the mixture was diluted with CH ₂ Cl ₂ (500 mL), washed with 5% citric acid (200 mL) and water, dewatered over Na 2 SO ₄ and evaporated to a small volume. The crystalline product was filtered and washed with CH 2 Cl 2 to give 4.14 g of the title compound. The mother liquors were dried and purified by flash chromatography (eluent: petrolatum / ethyl acetate 1: 1). The obtained solid was treated with petrolatum and filtered to give 1.9 g of the title compound which combined with the previous material gave a total yield of 6.04 g (yield: 73%).

¹ H-NMR (CDCl 3) δ: 10.60 (s-broad, 1H); 8.44 (d, 1H, J = 8Hz); 8.29 (s, 1 H); 7.49 (d, IH); 4.07 (s, 3H).

Example 60

6-Methoxy-5-phenyl-ethynyl-2H-phthalazin-1-one

A mixture of trifluoromethanesulfonic acid 6-methoxy-1-oxo-1,2-dihydrophthalazin-5-yl ester (1 g, 3.08 mmol), prepared as described in Example 59, phenylacetylene (0.507 mL, 4.62 mmol), bis- (triphenylphosphine) PdCl 2 (108 mg, 0.154 mmol), triethylamine (1.71 mL, 12.32 mmol) in DMF (20 mL) was heated at 90 ° C with stirring for 3 hours, then cooled, poured into water and extracted with CH 2 Cl 2. The extract was

- 55 washed with water, dehydrated over Na ₂ SO ₄ and dried. The residue was taken up in ethyl ether and filtered to give 680 mg of the title compound (yield: 80%).

¹ H-NMR (DMSO) δ: 12.70 (s-broad, 1H); 8.53 (s, 1 H); 8.25-7.45 (m. 7H); 4.03 (s, 3H).

Example 61

1-Chloro-6-methoxy-5-phenyl-ethynyl-phthalazine

A suspension of 6-methoxy-5-phenylethynyl-2H-phthalazin-1-one (0.67 g, 2.42 mmol), prepared as described in Example 60, in POCl 3 (20 mL) was heated to 80 ° C with stirring until dissolved. After 30 min, the solution was dried and the residue taken up in CH ₂ Cl ₂ (100 mL), washed to alkaline with K ₂ CO ₃ solution, then water, dewatered over Na ₂ SO ₄ and evaporated to give 715 mg of the title compound. compounds (quantitative yield). The compound was used as such in the next step.

¹ H-NMR (CDCl ₃ ) δ: 10.17 (s-broad, 1H); 8.51 (d, 1H, J = 10Hz); 8.07 (d, IH); 8.70-7.37 (m, 6H); 4.25 (s, 3H).

Example 62

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-5-phenyl-ethynyl-phthalazine (Compound 30)

To a solution of 3,5-dichloro-4-methylpyridine (0.768 g, 4.74 mmol) under dry N ₂ in dry DMF (10 mL) was added 60% NaH (189.6 mg, 4.74 mmol) at room temperature. and the mixture was stirred for 1 h, then 1-chloro-6-methoxy-5-phenylethynyl-phthalazine (700 mg, 2.37 mmol), prepared as described in Example 61, in dry DMF (20 mL) was added dropwise. . After 3 hours, the mixture was poured into water, extracted with ethyl acetate, washed with water, and dried over Na ₂ SO ₄

- 56 φφ φφ ♦ 9 9 Φ

Φ Φ Φ • Φ Φ ΦΦΦ

ΦΦΦΦ ΦΦΦΦ and dried. The residue was purified by flash chromatography (eluent: petrolatum / ethyl acetate 37). The isolated product was treated with ethyl ether and crystallized from 30 ml of ethyl acetate / _CH2 Cl2 to give 600 mg of the title compound (yield: 60%). Melting point: 217-219 ° C ¹ H-NMR (CDCl 3) δ: 9.88 (s, 1H); 8.51 (s, 2 H); 8.20-7.38 (m, 7H); 4.90 (s, 2 H); 4.13 (s, 3H).

Example 63

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-5-styryl-phthalazine (Compound 31)

A solution of 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-5-phenyl-ethynyl-phthalazine (0.47 g, 1.12 mmol), prepared as described in Example 62, in THF (50 mL) was hydrogenated in a Parr apparatus in the presence of 10% Pd / C (0.1 g) for 2 hours, then filtered, dried and purified by flash chromatography (eluent: petrolatum / ethyl acetate 1: 1). Product containing fractions were concentrated and treated with 20 mL of ethyl ether / petrolatum 1: 1. The crystallized compound was filtered and dried under vacuum at 40 ° C to give 0.37 g of the title compound (yield: 78%). Melting point: 180-182 ° C.

¹ H-NMR (CDCl 3) δ: 9.37 (s, 1H); 8.48 (s, 2 H); 8.17 (d, 1H, J = 9.2Hz); 7.62 (d, IH); 7.07 - 6.65 (m, 7H); 4.89 (s, 2 H); 3.92 (s, 3H).

Example 64

6-Methoxy-5- (5-phenylpent-1-ynyl) -2H-phthalazin-1-one

A mixture placed under N ₂ 6-methoxy-1-oxo-1,2-dihydrophthalazin-5-yl trifluoromethanesulfonic acid ester (2 g, 6.16 mmol), prepared as described in Example 59, pent-4-ynyl-benzene (1.33 g, 9.24 mmol), bis (triphenylphosphine) PdCl ₂ (216 mg, 0.308 mmol) and triethylamine (3.42 mL, 9 mL).

57

9 9

9 • 9 9 9

9

9999 999 • 9 9 9 9 • 99 999

24.64 mmol) in DMF (40 mL) was heated at 90 ° C for 3 h, then poured into water (200 mL), filtered and the precipitate was washed with water to give 1.28 g of the title compound (yield: 65%). .

¹ H-NMR (DMSO) δ: 12.63 (s, 1H); 8.41 (s, 1 H); 8.16 (d, 1H, J = 8.8Hz); 7.6 (d, H); 7.34-7.14 (m, 5H); 3.98 (s, 3H); 2.82-2.53 (m, 4H); 1.98-1.83 (m)

Example 65

1-Chloro-6-methoxy-5- (5-phenylpent-1-ynyl) -phthalazine

A suspension of 6-methoxy-5- (5-phenylpent-1-ynyl) -2H-phthalazin-1-one (1.26 g, 3.96 mmol), prepared as described in Example 64, and POCl 3 (30 mL) was heated to 80 ° C with stirring until dissolved. After 30 min, the solution was dried, the residue was dissolved in CH 2 Cl 2, washed to alkalinity with dilute K ₂ CO ₃ solution, then water, dehydrated over Na 2 SO ₄ and dried to give 1.33 g of the title compound (quantitative yield). The compound was used as such in the next step.

¹ H-NMR (CDCl ₃ ) δ: 9.77 (s, 1H); 8.21 (d, 1H, J = 9.2Hz); 7.59 (d, IH); 7.34-7.14 (m, 5H); 4.08 (s, 3H); 2.89-2.58 (m, 4H); 2.10-1.96 (m).

Example 66

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-5- (5-phenylpent-1-ynyl) phthalazine (Compound 32)

A solution of N _{2 of} 3,5-dichloro-4-methylpyridine (1.28 g, 7.9 mmol) in dry DMF (15 mL) was mixed with 60% NaH (316 mg, 7.9 mmol) with stirring at room temperature. ). The mixture was stirred for 1 h, then 1-chloro-6-methoxy-5- (5-phenylpent-1-ynyl) -phthalazine (1.33 g, 3.95 mmol), prepared as described in the example, was added dropwise. 65, in dry DMF (20 mL). After 3 h, the mixture was poured into water, extracted

- 58 • · «· ····

4 ·· «

4 4 · 8 · ·

9 4 9 9 9 with ethyl acetate, washed with water, dehydrated over Na ₂ SO ₄ and dried. The residue was purified by flash chromatography (eluent: petrolatum / ethyl acetate 1 L). The separated product was treated in ethanol (10 mL) and crystallized. The obtained product was filtered and dried under vacuum at 40 ° C to give 940 mg of the title compound (yield: 51%). Melting point: 130 - 132 ° C.

¹ H-NMR (CDCl 3) δ: 9.78 (s, 1H); 8.50 (s, 2 H); 8.12 and 7.60 (2d, 2H); 7.33-7.14 (m, 5H); 4.88 (s, 2 H); 4.09 (s, 3H); 2.89-2.81 (m, 2 H); 2.62 (t, 2H, J = 7.0Hz); 2.11-1.96 (m, 2 H).

Example 67

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-5- (5-phenyl-pent-1-enyl) -phthalazine (Compound 33)

A solution of 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-5- (5-phenyl-pent-1-ynyl) -phthalazine (280 mg, 0.6 mmol), prepared as described in the Example 66, in THF (15 mL) was hydrogenated in a Parr apparatus in the presence of 10% Pd / C (50 mg) for 1 h, then filtered, dried and purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH ₃ OH 98: 2 ). The separated oil was taken up in ethyl ether (5 mL) and crystallized. The crystalline obtained material was filtered and dried under vacuum at 40 ° C to give 120 mg of the title compound (yield: 43%). Melting point: 121 - 123 ° C.

¹ H-NMR (CDCl ₃ ) δ: 9.42 (s, 1H); 8.51 (s, 2 H); 8.16 and 7.62 (2d, 2H, J = 9.1 Hz); 7.20-6.97 (m, 5H); 6.50 (d-broad, 1H, J = 11.4 Hz); 6.14 (dt, 1H, J = 7.1Hz); 4.90 (s, 2 H); 3.99 (s, 3H); 2.51-2.43 (m, 2 H); 1.941.63 (m, 4H).

- 59 99 ·· ·· • 9 9 9 9 • * · 9 · 9 9 9

9 9 9 • 9 9999 99

Example 68

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-5- (5-phenyl-pentyl-D-phthalazine hydrochloride) (Compound 34)

A solution of 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-5- (5-phenyl-pent-1-ynyl) -phthalazine (770 mg, 1.655 mmol), prepared as described in Example 66, in THF (50 mL) was hydrogenated in a Parr apparatus in the presence of 10% Pd / C (100 mg) for 24 h. The solution was dried and purified by flash chromatography (eluent: petrolatum / ethyl acetate 1 L). The fractions corresponding to the starting compound were dried and hydrogenated again for 24 hours and then worked up as above. Fractions containing the desired product were combined with the fractions obtained above, dissolved in ethyl acetate, and acidified with HCl / ethyl acetate. The precipitate was filtered off and crystallized from ethanol (2 ml) and ethyl acetate (8 ml) followed by concentration. 142 mg of the title compound were obtained (yield: 17%). Melting point: 170-173 ° C.

¹ H-NMR (CDCl 3) δ: 9.83 (s, 1H); 8.54 (s, 2 H); 8.39 and 7.97 (2d, 2H, J = 9.1 Hz); 7.29-7.11 (m, 5H); 5.02 (s, 2 H); 4.12 (s. 3H); 3.16-3.08 and 2.63-2.56 (2m, 4H); 1.72-1.38 (m, 6H).

Example 69

5-Benzyloxy-6-methoxy-2H-phthalazin-1-one

Glacial acetic acid (285 mL) at 10-20 ° C was mixed with hydrazine monohydrate (10.16 mL, 209 mmol) added dropwise and 3-benzyloxy-N, N -diethyl-2-formyl-4 was added. -methoxy-benzamide (14.27 g, 41.8 mmol), prepared as described in Example 56. The mixture was allowed to stand for 2 hours, then evaporated and dissolved in CH 2 Cl 2, washed with water, dehydrated over Na ₂ SO 4, dried and the crude material was triturated in petrolatum / ethyl ether

-60 ·· ··. 9 ♦ 9 1 1 (150 ml) 9 9 9: 1 (150 ml) ) and filtered to give 11.3 g of the title compound (yield: 96%).

¹ H-NMR (CDCl 3) δ: 10.61 (s, 1H); 8.30 (s, 1 H); 8.18-7.31 (m, 7H); 5.17 (s, 2 H); 4.03 (s, 3H).

Example 70

5-Benzyloxy-1-chloro-6-methoxyphthalazine

A suspension of 5-benzyloxy-6-methoxy-2H-phthalazin-1-one (4 g, 14.17 mmol), prepared as described in Example 69, in acetonitrile (40 mL) and POCl 3 (6.5) mL, 70.8 mmol) was heated to 80 ° C with stirring until dissolved. After 30 min, the solution was evaporated and the residue taken up in CH 2 Cl 2 (100 mL), washed with cold NaOH solution (20 mL) and water (200 mL) and extracted into ethyl ether (twice) and ethyl acetate (once) to check that the pH around seven. The organic phase was washed with saturated NaCl solution, dewatered and concentrated to give 4.15 g of the title compound (yield: 97%).

¹ H-NMR (CDCl 3) δ: 9.49 (s, 1H); 7.95 (d, 1H, J = 9.0Hz); 7.62 (d, IH); 7.41-7.28 (m, 5H); 5.24 (s, 2 H); 4.86 (s, 2 H); 4.06 (s, 3H).

Example 71

5-Benzyloxy-1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazine

A solution of 3,5-dichloro-4-methylpyridine (5.57 g, 34.38 mmol) in DMF (30 mL) under N ₂ was treated with NaH (60% in oil, 1.38 g, 34.38 mmol) and the mixture was heated in a water bath at 40 ° C. After 1 h, a solution of 5-benzyloxy-1-chloro-6-methoxyphthalazine (4.15 g, 13.75 mmol), prepared as described in Example 70, in DMF (30 mL) was added dropwise and the whole was allowed to stand. at room temperature for 2 hours and then poured into brine and extracted with ethyl acetate. Earned

- 61 • 999 • 9 9 9 • 9 9

9 9

9 9

9999

9 9 9

9

9999999 9 • 9 9 9 9 was poured into brine and pH = 7 buffer and extracted three times with ethyl acetate. The solid obtained was filtered, allowed to air dry, dissolved in CH ₂ Cl ₂ and concentrated. The organic phase was dewatered and concentrated to give a solid which, trituration in ethyl acetate + propyl ether gave 3.866 g, which combined with the solids treated above yielded a total of 4.346 g of the title compound (yield: 78%).

¹ H-NMR (CDCl 3) δ: 9.60 (s, 1H); 8.49 (s, 2 H); 7.93 (d, 1H, J = 9.2Hz); 7.66 (d, IH); 7.49-7.31 (m, 5H); 5.26 (s, 2 H); 4.86 (s, 2 H); 4.08 (s, 3H).

Example 72

1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazin-5-ol dihydrochloride

5-Benzyloxy-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (4.3 g, 10.08 mmol), prepared as described in Example 71, in glacial acetic acid and 37% HCl was placed in a flask with stirring at room temperature for 1/2 hour and then at 60 ° C for 1 hour. The mixture was dried, treated several times with toluene to give a solid which was triturated in acetone and dried to give 4.04 g of the title compound (yield: 98%).

¹ H-NMR (DMSO) δ: 10.14 (s, 1H); 8.72 (s, 2 H); 8.35-8.25 (m, 2 H); 5.10 (s. 2H); 4.12 (s, 3H).

Example 73

Trifluoromethanesulfonic acid - (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl ester

Suspension under N _{2 of} 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazin-5-ol (0.469 g, 1.4 mmol), prepared as described ♦ 9 · * ·· · ·· · · · · · · · · »» »» »» »» »» »»

Example 72, in acetonitrile (5 mL) at 40 ° C was mixed with K ₂ CO ₃ (0.77 g, 5.58 mmol), a catalytic amount of Na ₂ S ₂ O _5, and N, N-bis (trifluoromethanesulfonyl) phenylamine (1 g, 2.8 mmol). The suspension turned red and turned into a solution that was dried, the residue taken up in CH 2 Cl 2 and washed with 5% citric acid and water. The solution was dewatered and concentrated to give a solid which was triturated in ethyl ether to give 1 g of the title compound (yield: 56%).

¹ H-NMR (CDCl 3) δ: 9.52 (s, 1H); 8.41 (s, 2 H); 8.27 (d, 1H, J = 9.0Hz); 7.79 (d, IH); 5.26 (s, 2 H); 4.87 (s, 2 H); 4.0.8 (s. 3H).

Example 74

Benzyl- {3-ri- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl-prop-2-invyl) -methylamine dihydrochloride (Compound 35)

To a suspension under _N2 of 1- (3,5-dichloro-4-ylmethyl) -6-methoxy-phthalazin-5-yl ester (5 g, 10.68 mmol), prepared as described in Example 73, 2-benzylmethylprop -inyl-amine (2.16 mL, 12.82 mmol) and diethylamine (100 mL) were added with stirring bis (triphenylphosphine) PdCl ₂ (150.2 mg, 0.214 mmol) and CuI (40.75 mg, 0.214 mmol) ). The mixture was refluxed for 6 hours and the residue was purified by flash chromatography (eluent: ethyl acetate) to give a solid which was dissolved in CH 2 Cl 2 / CH 3 OH 1: 1 (50 mL), acidified with HCl in ethyl acetate and dried. The residue was dissolved in CH ₂ Cl ₂ (100 mL), dried again, triturated in ethyl acetate (150 mL), filtered and dried in an oven under vacuum at 40 ° C to give 5.22 g of the title compound (yield: 83%). ).

¹ H-NMR (DMSO) δ: 11.70 (s-broad, 1H); 9.91 (s, 1 H); 8.72 and 8.13 (2d, 2H, J = 9.3 Hz); 8.69 (s, 2 H); 7.69-7.45 (m, 5H); 5.05 (s, 2 H); 4.54-4.37 (m, 4H); 4.15 (s. 3H); 2.48 (s, 3H).

-63 • 9 • 9

9 9 • 999 99

99999 * 9 9 • 9 9 9 9

9 ··

Example 75

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-5- (5-morpholin-5-yl-pent-1-ynyl) phthalazine dihydrochloride (Compound 36)

Trifluoromethanesulfonic acid 1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl ester (0.935 g, 2 mmol), prepared as described in Example 73, 4-pent-4-ynylmorpholine (O, 37 g, 2.4 mmol), diethylamine (9 mL) and dry acetonitrile (6 mL), then bis (triphenylphosphine) PdCl ₂ (0.014 g, 0.02 mmol) and CuI (0.004 g, 0.02 mmol) ) were placed in a flask under N ₂ and the mixture was stirred at room temperature overnight, dried, the residue taken up in ethyl acetate and washed with water. The organic phase was washed with water, dewatered and concentrated to give an oil which was purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH 3 OH / NH ₃ 95: 5: 0.5) to give an oil which was dissolved in ethyl acetate and precipitated by addition of HCl in ethyl ether. 0.09 g of the title compound was obtained. Melting point: 129 - 131 ° C (dec.).

¹ H-NMR (DMSO) δ: 9.71 (s, 1H); 8.67 (s, 2 H); 8.59 and 8.02 (2d, 2H, J = 9.2 Hz); 5.00 (s, 2 H); 4.08 (s, 3H); 4.02-3.63 (m, 4H); 3.49-2.70 (m, 8H); 2.14-1.99 (m, 2 H).

Example 76

3-ri- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxyphthalazin-5-yl-prop-2-yn-1-ol (compound 37)

Under N ₂ were charged to a flask prop-2-in-1-ol (0.151 mL, 2.55 mmol), triethylamine (0.95 mL, 6.8 mmol), 1- (3,5-dichloropyridin-4- trifluoromethanesulfonic acid-6-methoxyphthalazin-5-yl ester (0.795 g, 1.7 mmol), prepared as described in Example 73, bis (triphenylphosphine) PdCl ₂ (0.06 g, 0.08 mmol) and DMF (10 mL) and heated at 85 ° C for 5 h, then the mixture was maintained at temperature

9

9

- 64 9 • 9 · 9 • 9 · 9

9 9

9 9

9 9

9999 ·· 9

9 9

9 9 9

99999

9 «

9

9

9

9

9 laboratories overnight. The mixture was diluted with ethyl acetate and washed with water. The organic phase was dewatered and concentrated to give a solid which was triturated with ethyl ether. Addition of this product to the product obtained by extraction and chromatography of the mother liquors (eluent; CH 2 Cl 2 / CH 3 OH / NH 3 95: 5: 0.5) afforded 0.164 g of the title compound (yield: 26%).

¹ H-NMR (DMSO) δ: 9.69 (s, 1H); 8.68 (s, 2 H); 8.57 (d, 1H, J = 9.3Hz); 7.79 (d, IH); 5.50 (t, IH); 4.98 (s, 2 H); 4.47 (d, 2H, J = 5.9 Hz); 4.08 (s, 3H).

Example 77

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4-morpholin-4-yl-phthalazine (Compound 38)

To a solution of 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazine (1 g, 2.82 mmol), prepared as described in Example 45, in DMF (25 mL), at room temperature, morpholine (0.73 g, 8.46 mmol) was added under stirring and under a dry N ₂ atmosphere and the temperature was raised to 100 ° C. After 14 h, all the mixture was dried and the solid was partitioned between water and CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and dried to give a solid which was purified by flash chromatography (eluent: hexane / ethyl acetate 6: 4 then in a gradient up to 3: 7) to give 0.9 g of the title compound (yield). : 79%). Melting point: 179-180 ° C.

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 8.10-7.34 (m, 3H); 4.78 (s, 2 H); 3.98 (s, 3H); 3.96-3.91 (m, 4H); 3.41-3.36 (m, 4H).

φφ φ

-65 ·· φ φ • φ • φ φ φ φ

«

Example 78 4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-phthalazin-1-yl-propylamine (Compound 39)

4-Chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazine (3.9 g, 11 mmol), prepared as described in Example 45, and n-propylamine (40 mL) ) were placed in an autoclave at an external temperature of 120 ° C (internal temperature 100 ° C, 0.4 MPa) for 2 hours. The solution was concentrated to give a residue, taken up in CH ₂ Cl ₂ and 5% HCl and extracted several times. The aqueous phase was basified with K ₂ CO ₃ and extracted with CHOCl ₂ . The obtained solid was crystallized from acetonitrile (110 mL) and then filtered to give 3.26 g of the title compound (yield: 82%). M.p .: 222-223 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.42 (s, 2H); 7.99 (d, 1H, J = 9.0Hz); 7.44 (dd, IH); 7.03 (d, 1H, J = 2.5Hz); 4.89 (t, 1H, J = 5.3 Hz); 4.68 (s, 2 H); 3.95 (s, 3H); 3.63-3.53 (m, 2 H); 1.79-1.60 (m, 2 H); 0.96 (t, 3H, J = 7.3Hz).

Example 79

4- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxy-1 H -phthalazine-2-carboxylic acid hydroxyamide (Compound 40)

At room temperature, with stirring under N ₂ , to a solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazine (1 g, 3.1 mmol), prepared as described in Example 22, in THF (35 mL), 1,1'-carbonyldiimidazole (0.55 g, 3.41 mmol) was added and the solution was heated to reflux, concentrated and taken up in ethanol. Hydroxylamine hydrochloride (0.26 g, 3.72 mmol) in ethanol was added. After 10 h under reflux, the mixture was cooled, concentrated, and partitioned between water and CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and dried to give a residue which was purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH ₃ OH 98: 2, then hexane / ethyl acetate).

-66> * * ·· • · · · · · · Ι · Ι · Ι · »· * · · · · · * · · · M + H: 7), yielding 0.3 g of the title compound (yield: 25%). M.p .: 155-158 ° C.

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 7.72 (d, 1H, J = 3.5Hz); 7.45 (d, 1H, J = 8.7Hz); 7.22 (d, IH); 6.88 (dd, IH); 6.69 (d, 1H, J = 2.5Hz); 4.79 (s, 2 H); 4.27 (s, 2 H); 3.84 (s, 3H).

Example 80

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1H-phthalazin-2-yl-oxo-acetic acid ethyl ester (Compound 41)

At room temperature, under N ₂ and with stirring, to a solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazine (2 g, 6.2 mmol), prepared as described in of Example 22, in THF (70 mL), triethylamine (2.16 mL, 15.5 mmol) was added, the solution was cooled to 0 ° C, and ethyloxalyl chloride (0.83 mL, 7.44 mmol) was added dropwise. After 30 min, cooling was stopped and after 5 h at room temperature the mixture was poured into water / ice, concentrated and extracted several times with CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and dried to give a solid which crystallized from acetonitrile to give 1.87 g of the title compound (yield: 73%). Melting point: 140-142 ° C.

¹ H-NMR (CDCl 3) δ: 8.46 (s, 2H); 7.52-6.70 (m, 3H); 4.87 (s, 2 H); 4.28 (s, 2 H); 3.86 (s, 3H); 3.79 (q, 2 H); 1.15 (t, 3H, J = 7.1Hz).

Example 81

2-Chloro-1-F4- (3,5-dichloropyridin-4-ylmethyl) -7-methyl) -7-methoxy-1 H -phthalazin-2-yl-ethanone

At room temperature, under N ₂ and with stirring, to a solution of 1- [4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazine (1.9 g, 5.9 mmol), prepared as was described in Example 22, in dry THF

- 67 99 ·· • 9 9 • · 99 999 999

9999 99 9 99 999 (70 mL) was added triethylamine (2.05 mL, 14.7 mmol) and the mixture was cooled to 0 ° C. Chloroacetyl chloride (0.56 mL, 7.07 mmol) was added dropwise and a precipitate formed. After 1 night an additional 50% triethylamine and chloroacetyl chloride was added, and after 20 hours at room temperature, the mixture was poured into water / ice, concentrated and extracted several times with CH 2 Cl 2. The organic phase was washed with water, dewatered and dried to give 2.6 g of the title compound (quantitative yield).

Example 82

1- [4- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazin-2-yl-2-morpholin-4-yl-ethanone (Compound 42)

To a solution under N ₂ , and with stirring, 2-chloro-1- [4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazin-2-yl] -ethanone (1,3 g, 3.26 mmol), prepared as described in Example 81, in dry SHCl ₃ (15 mL) at room temperature, morpholine (0.85 g, 9.78 mmol) was added and the mixture was refluxed. The mixture was poured into water / ice, the phases were separated and the organic phase was washed with water, dewatered and dried to give a solid that crystallized from acetonitrile (45 mL) to give 0.77 g of the title compound ( yield: 53%). Melting point: 180-182 ° C.

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 7.46 (d, 1H, J = 8.5Hz); 6.89 (dd, IH); 6.71 (d, 1H, J = 2.6Hz); 4.85 (s, 2 H); 4.29 (s, 2 H); 3.85 (s, 3H); 3.70-3.65 (m, 4H); 3.01 (s, 2 H); 2.39-2.35 (m, 4H).

Example 83

1- [4- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazin-2-yl] -2-pyrrolidin-1-yl-ethanone (Compound 43)

To a solution under N ₂ , and with stirring, 2-chloro-1- [4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1 H -phthalazin-2-yl] -ethanone (1, 2-chloro) 3 g, 3.26 mmol), prepared as described in Example 81, in dry CHCl ₃ (15 mL) at room temperature, pyrrolidine (0.82 mL, 9.78 mmol) was added and the mixture was refluxed. . After 20 h, it was poured into water / ice, the phases were separated and the organic phase was washed with water, dewatered and dried to give a residue which crystallized from acetonitrile (40 mL) to give 0.79 g of the title compound (yield: 58%). M.p .: 148-150 ° C.

¹ H-NMR (CDCl 3) δ: 8.49 (s, 2H); 7.45 (d, 1H, J = 8.6Hz); 6.88 (dd, 1 H, J); 6.70 (d, 1H, J = 2.6Hz); 4.85 (s, 2 H); 4.28 (s, 2 H); 3.84 (s, 3H); 3.14 (s, 2 H); 2.48-2.41 (m, 4H); 1.77-1.68 (m, 4H).

Example 84 [4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-phthalazin-1-yl-dimethyl-amine (Compound 44)

At room temperature, a solution of [4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxyphthalazin-1-yl] -propylamine (3.5 g, 5.64 mmol) was prepared as described in the Example. 78, in DMF (35 mL), stirred under N ₂ , K ₂ CO ₃ (0.78 g, 5.64 mmol) and methyl iodide (0.35 mL, 5.64 mmol) were added. The mixture was heated to 50 ° C and after 20 hours dried and partitioned between water and CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and concentrated to give an oil which was purified by flash chromatography (eluent: hexane / ethyl acetate 4: 6) to give 1 g of the title compound (yield: 50%). Melting point: 170-173 ° C.

¹ H-NMR (CDCl 3) δ: 8.46 (s, 2H); 8.05-7.39 (m, 3H); 4.75 (s, 2 H); 3.98 (s, 3H); 3.07 (s, 6H).

• · · · ······ ·

- 69 Example 85

1- (3,5-Dichloro-pyridin-4-ylmethyl) -4-imidazol-1-yl-6-methoxy-phthalazine (Compound 45)

A solution of imidazole (0.814 g, 11.21 mmol) in dry DMF (30 mL) was stirred under N ₂ at room temperature and NaH (8.46 mmol) was added. After 30 min, 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (2 g, 5.64 mmol), prepared as described in Example 45, in dry DMF (30 mL) was added. ). The mixture was heated at 80 ° C for 3.5 hours, cooled, poured into water / ice and extracted three times with CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and concentrated to give a solid which was purified by flash chromatography (eluent: ethyl acetate) to give 0.9 g of the title compound (yield: 50%). Melting point: > 250 ° C.

¹ H-NMR (CDCl 3) δ: 8.53 (s, 2H); 8.27-7.31 (m, 6H); 4.94 (s, 2 H); 3.95 (s, 3H).

Example 86: 4- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxyphthalazin-1-yl-thiazol-2-yl-amine (Compound 46)

A solution of 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (0.5 g, 1.41 mmol), prepared as described in Example 45, 2-, was stirred at 100 ° C. aminothiazole (0.424 g, 4.23 mmol), triethylamine (0.59 mL, 4.23 mmol), palladium acetate (16 mg, 0.07 mmol) and triphenylphosphine (56 mg, 0.21 mmol) under N ₂ . After 20 h, the mixture was poured into water / ice and extracted several times with CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and concentrated to give a solid which was purified by flash chromatography (eluent: hexane / ethyl acetate 7: 3) to give 0.24 g of the title compound (yield: 50%). Melting point: 218-220 ° C.

¹ H-NMR (CDCl 3) δ: 14.41 (s, 1H); 8.51 (s, 2 H); 8.09 (d, 1H).

- 70 - ······ * .. *

J = 2.7 Hz); 7.81 (d, 1H, J = 8.8Hz); 7.42-7.36 (m, 2 H); 6.82 (d, 1H, J = 3.8Hz); 4.55 (s, 2 H); 4.03 (s, 3H).

Example 87

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-4-phenoxyphthalazine (Compound 47)

A suspension of DMF (15 mL), phenol (0.53 g, 5.64 mmol) and NaH (4.23 mmol) was stirred under dry N ₂ at room temperature. After 15 min, 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (0.5 g, 1.41 mmol), prepared as described in Example 45, was added dropwise and the temperature was brought at 100 ° C. After 20 h, the mixture was partitioned between water and CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and concentrated to give a residue which was triturated in ethyl ether. The insoluble material was purified by flash chromatography (eluent: hexane / ethyl acetate 7: 3) to give a solid that crystallized from isopropyl ether (15 mL) to give 0.25 g of the title compound (yield: 49%). Melting point: 130 - 132 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.43 (s, 2H); 8.10-7.54 (m, 3H); 7.43-7.15 (m, 5H); 4.78 (s, 2 H); 4.03 (s, 3H).

Example 88

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-4- (4-methylpiperazin-1-yl) phthalazine (Compound 48)

A solution of 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazine (1 g, 2.82 mmol), prepared as described in Example 45, in DMF (25 mL) was stirred under dry N ₂ at room temperature, and 1-methylpiperazine (0.848 g, 8.46 mmol) was added dropwise. The mixture was heated at 100 ° C for 20 h, then concentrated to small volume and partitioned between water CH ₂ Cl ₂ . The organic phase was

washed with water, dewatered and concentrated to give a residue which was purified by flash chromatography (eluent: CH 2 Cl 2 / CH 3 OH / NH 4 OH 95: 5: 0.5). The obtained solid was crystallized from tert-butyl methyl ether (35 mL) to give 0.4 g of the title compound (yield: 34%). Melting point: 160-162 ° C.

¹ H-NMR (CDCl 3) δ: 8.47 (s, 2H); 8.05 (d, 1H, J = 9.1Hz); 7.42 (dd, IH); 7.36 (d, 1H, J = 2.5Hz); 4.77 (s, 2 H); 3.98 (s, 3H); 3.49-3.44 (m, 4H); 2.69-2.63 (m, 4H); 2.37 (s, 3H).

Example 89

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-4-pyrrolidin-1-yl-phthalazine (Compound 49)

A solution of 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazine (1 g, 2.82 mmol), prepared as described in Example 45, in DMF (25 mL) was stirred under dry N ₂ at room temperature, and pyrrolidine was added dropwise. The mixture was heated at 100 ° C for 20 h, then was concentrated and partitioned between water and CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and concentrated to give a solid that crystallized from ethyl acetate (25 mL) to give 0.77 g of the title compound (yield: 71%). Melting point: 170-172 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.45 (s, 2H); 7.98 (d, 1H, J = 9.1Hz); 7.55 (d, 1H, J = 2.6Hz); 7.44 (dd, IH); 4.69 (s, 2 H); 3.95 (s, 3H); 3.85-3.78 (m, 4H); 2.00-1.94 (m, 4H).

Example 90

[4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-1-oxo-1 H -phthalazin-2-yl] -acetic acid (Compound 50)

To a suspension of [4- (3,5-dichloro-pyridin-4-yl) -ethyl ester •

- 72-methyl) -7-methoxy-1-oxo-1H-phthalazin-2-yl] -acetic acid (0.65 g, 1.54 mmol), prepared as described in Example 44, in ethanol (30 mL) ), concentrated NaOH (3 mL) was added. The mixture was stirred overnight at room temperature, then dried, taken up in water and washed twice with CH 2 Cl 2. The aqueous phase was acidified with concentrated HCl. The precipitate was filtered and dried over P ₂ O ₅ at 60 ° C, then crystallized from CH 3 OH (25 mL) and the filtrate was dried at 45 ° C to give 0.36 g of the title compound (yield: 60%). Melting point: 221.4 - 222.4 ° C.

¹ H-NMR (CDCl 3) δ: 8.63 (s, 2H); 8.20 (d, 1H, J = 8.8Hz); 7.68 (d, 1H, J = 2.8Hz); 7.60 (dd, IH); 4.61 and 4.50 (2s, 4H); 3.96 (s, 3H).

Example 91

2-14- (3,5-Dichloropyridin-4-ylmethyl) -7-methoxy-1-oxo-1 H -phthalazin-2 H-N-hydroxyacetamide (Compound 51)

To a solution of [4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1-oxo-1 H -phthalazin-2-yl] -acetic acid (0.61 g, 1.55 mmol), prepared as described in Example 90, in dry DMF (25 mL), under N 2, 1,1'-carbonyldiimidazole (0.38 g, 2.32 mmol) was added. The mixture was stirred at room temperature for 4 hours, then cooled, hydroxylamine hydrochloride (0.18 g, 2.55 mmol) was added and the mixture was stirred overnight. A precipitate formed and the solution was dried in vacuo and then taken up in 10% NaOH. Stirring was continued for 2 hours, then the solid was filtered, washed with water, then with acetone and ethyl ether. This material was taken up in 5% HCl and stirred for 1 hour, then the mixture was filtered and washed with water, acetone and ethyl ether. The solid was dried under vacuum at 50 ° C, crystallized from acetic acid (60 mL) and dried under vacuum at 40 ° C to give 0.3 g of the title compound (yield: 40%). Melting point: 256.4-258 ° C.

¹ H-NMR (DMSO) δ: 10.51 (broad, 1H); 8.82 (broad, 1H); 8.63 (s,

2H); 8.21-7.57 (m, 3H); 4.60 (s, 2 H); 4.35 (s, 2 H); 3.96 (s, 3H).

Example 92

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4- [1,2,41-triazol-1-yl-phthalazine-3-oxide (compound 52)

To a solution under N ₂ at 0 ° C 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-4- [1,2,4] triazol-1-yl-phthalazine (5.1 g, 13, 2 mmol), prepared as described in Example 51, in trifluoroacetic acid (25 mL), H ₂ O ₂ (1.35 mL, 13.5 mmol) was added dropwise. The temperature was allowed to rise to room temperature and after 48 hours the pH was adjusted with NaOH. After an additional 24 hours with stirring, the mixture was dried, taken up in CH ₂ Cl ₂ . The organic phase was washed with water, dewatered and dried to give a foam which was purified by flash chromatography (eluent: ethyl acetate / ethyl ether 6: 4 then 8: 2) to give 2.2 g of the title compound (yield: 41%). Melting point: 218-220 ° C (dec.).

¹ H-NMR (CDCl 3) δ: 9.12 and 8.26 (2s, 2H); 8.42 (s, 2 H); 8.48-7.49 (m, 3H); 4.90 (s, 2 H); 3.98 (s, 3H).

Example 93

6-Hydroxy-3H-isobenzofuran-1-one

To a solution of 6-methoxy-3/7-isobenzofuran-1-one (50 g, 0.3 mol), prepared as described in Example 34, in CH ₂ Cl ₂ (250 mL), under N ₂ at -5 ° C, BBr ₃ (360 mL, 0.36 mol) was added dropwise. After the addition was complete, the solution was stirred at room temperature overnight, then cooled and additional BBr ₃ (60 mL, 60 mmol) was added. Stirring was maintained for 4 hours at room temperature. The mixture was cooled and CH ₃ OH (250 mL) was added, then dried under vacuum to give a solid which was mixed with ethyl ether and washed with ethyl ether and ethyl acetate. After drying under vacuum at 40 ° C, a solid was obtained which was combined with the solid material obtained by drying the mother liquors, decolorizing and triturating in ethyl ether. 33.51 g of the title compound were obtained (yield: 73%).

¹ H-NMR (DMSO) δ: 10.05 (s, 1H); 7.50-7.08 (m, 3H); 5.29 (s,

2H).

Example 94

6-Difluoromethoxy-3/7-isobenzofuran-1-one

In an autoclave, to a solution of 6-hydroxy-3/7-isobenzofuran-1-one (34.06 g, 0.227 mol), prepared as described in Example 93, in DMF (340 mL) was added K 2 CO 3 (34.5 g). g, 0.25 mol) under freon atmosphere. After one night with vigorous stirring at 110 ° C, the mixture was cooled and K 2 CO 3 was filtered off. The solution was dried to give a solid which was taken up in water / ethyl acetate. Insoluble matter was filtered off and mixed with water and ethyl acetate. The organic phase was drained and dried under vacuum to give an oil which was taken up in CHCl 3. The solution was purified by flash chromatography (eluent: petrolatum / ethyl acetate 7: 3) to give 24.1 g of the title compound (yield: 53%).

¹ H-NMR (CDCl 3) δ: 7.61-7.40 (m, 3H); 6.56 (t, 1H, J = 72Hz); 5.29 (s, 2 H).

Example 95

3-Bromo-6-difluoromethoxy-3H-isobenzofuran-1-one

To a suspension of 6-difluoromethoxy-3 H -isobenzofuran-1-one (1 g, 5 mmol), prepared as described in Example 94, in CCl 4 (10 mL), under N ₂ , was added N-bromosuccinimide (0). (91 g, 5.1 mmol) and the mixture was heated at 76 ° C. A solution of α, cis -za-isobutyronitrile (0.01 g) in CHCl 3 (1 mL) was added slowly. The mixture was refluxed

1.5 hours, then cooled, filtered through celite and washed with CCl ₄ . After drying, 1.41 g of the title compound was obtained as an oil (quantitative yield).

¹ H-NMR (CDCl 3) δ: 7.65-7.49 (m, 3H); 7.37 (s, 1 H); 6.60 (t, 1 H, J = 72 Hz).

Example 96 (5-Difluoromethoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenyl-phosphonium bromide

Under N 2, to a solution of 3-bromo-6-difluoromethoxy-3H-iso-benzofuran-1-one (6.2 g, 22.2 mmol), prepared as described in Example 95, in dry acetonitrile (30 mL) was added triphenylphosphine (6.4 g, 24.4 mmol). After 4 hours at reflux, the mixture was dried to give a foam which was triturated in ethyl ether overnight, then filtered, washed with ethyl ether and dried to give 11.4 g of the title compound (yield: 95%).

¹ H-NMR (CDCl 3) δ: 9.96 (s, 1H); 7.89-7.16 (m, 18H); 6.58 (t, 1H, J _HF = 72 Hz).

Example 97

3- (3,5-Dichloro-pyridin-4-ylmethylene) -6-difluoromethoxy-3H-isobenzofuran-1-one

To a solution of (5-difluoromethoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenylphosphonium bromide (11.4 g, 21 mmol), prepared as described in Example 96, in dry CH 2 Cl 2 (110 mL) under N 2, 3,5-dichloro-4-formylpyridine (4.06 g, 23.1 mmol) was added. The mixture was cooled to 4 ° C and triethylamine (3.8 mL, 27.3 mmol) was added while maintaining the temperature below 10 ° C. After the addition was complete, the mixture was stirred

76 at room temperature for 3 hours, then cooled and 5% HCl was added. The organic phase was washed with aqueous NaCl solution, decolourised with charcoal, filtered through celite and dried, yielding 14.4 g of the title compound (stoichiometric yield containing 1 equivalents of triphenylphosphine oxide).

¹ H-NMR (CDCl 3) δ: 8.62 (s, 2H); 8.29-7.78 (m, 4H); 6.59 (t, 1H, J _HF = 72 Hz).

Example 98

4- (3 _l 5-Dichloro-pyridin-4-ylmethyl) -7-difluoromethoxy-2 / - / - phthalazin-1-one (Compound 53)

To a solution of 3- (3,5-dichloropyridin-4-ylmethylene) -6-difluoromethoxy-3H-isobenzofuran-1-one (14.4 g, 21 mmol), prepared as described in Example 97, in CH ₃ OH ( 100 mL), under N ₂ , acetic acid (3.6 mL, 63 mmol) and hydrazine monohydrate (3.15 mL, 63 mmol) were added. A precipitate formed and the mixture was refluxed for 2 hours. After standing overnight, the mixture was cooled with ice and the solid was filtered and washed with a small amount of CH ₃ OH. After drying in vacuo at 50 ° C, 6.64 g of the title compound were obtained [yield: 85% of (5-difluoromethoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenylphosphonium bromide].

¹ H-NMR (CDCl 3) δ: 12.54 (s, 1H); 8.65 (s. 2H); 8.36-7.80 (m, 3H); 7.58 (t, 1H, J _H = 74 Hz); 4.62 (s, 2 H).

Example 99

4-Chloro-1- (3 ₁ 5-dichloropyridine-4-ylmethyl) -6-difluormethoxvftalazin

To a suspension of 4- (3,5-dichloropyridin-4-ylmethyl) -7-difluoromethoxy-2H-phthalazin-1-one (6.64 g, 17.8 mmol), prepared as described in Example 98, in acetonitrile ( 70 mL), under N ₂ , was added POCl ₃ (83 mL, 9 mL)

- 77 • ·.

• 9 • 9 mmol). After 4.5 hours under reflux and 1 night standing, the mixture was dried under vacuum, taken up in CHOCl 3 and washed with aqueous Na 2 CO 3, decolourised with charcoal, filtered through celite and dried again to give 6.8 g of the title compound (yield: 97%).

¹ H-NMR (CDCl 3) δ: 8.50 (s, 2H); 8.29-7.78 (m, 3H); 6.77 (t, 1H, J H F = 72 Hz); 4.89 (s, 2 H).

Example 100

1- (3,5-Dichloropyridin-4-ylmethyl) -6-difluoromethoxy-4- [1,2,41-triazol-1-yl-phthalazine (Compound 54)

To a solution of 1,2,4-triazole under N ₂ (2.4 g, 34.8 mmol) in DMF (100 mL) was added NaH (1.05 g, 26.1 mmol). The mixture was stirred at room temperature until clear and then 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-difluoromethoxyphthalazine (6.8 g, 17.4 mmol), prepared as described, was added. in Example 99. The mixture was heated at 100 ° C for 5 hours, extracted three times with CH 2 Cl 2. The organic phase was dewatered and dried to give a solid which was purified by flash chromatography (eluent: ethyl acetate / petrolatum 7: 3 to 100% ethyl acetate) to give 4.14 g of the title compound (yield: 28%). Melting point: 220-224 ° C.

¹ H-NMR (CDCl 3) δ: 9.26 and 8.27 (2s, 2H); 8.97 (d, 1H, J = 2.4Hz); 8.55 (s. 2H); 8.36 (d, 1H, J = 7.2Hz); 7.86 (dd, IH); 6.79 (t, 1H, J = 72Hz); 4.97 (s, 2 H).

Example 101

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-difluoromethoxy-4-morpholin-4-yl-phthalazine (Compound 55)

Suspension under N _{2 of} 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-difluoromethoxyphthalazine (5.5 g, 14.1 mmol), prepared as described in Example 99, in morpholine ( 11 mL) was heated at 100 ° C for 1 h, then cooled and poured into water (150 mL). The solid that formed was stirred for 15 min, filtered, washed with water, dried over P ₂ O ₅ at 50 ° C under high vacuum. 6 g of the title compound were obtained (yield: 96%). Melting point: 128.6 130.6 ° C.

¹ H-NMR (CDCl ₃ ) δ: 8.49 (s, 2H); 8.20-7.65 (m, 3H); 6.70 (t, 1H, J = 71.9Hz); 4.81 (s, 2 H); 3.95-3.91 (m, 4H); 3.47-3.42 (m, 4H).

Example 102

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-phthalazine-1-carboxylic acid methyl ester

To a suspension of 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazine (9.7 g, 27 mmol), prepared as described in Example 45, in DMSO / CH 3 OH (80 / 40 mL) was added K ₂ CO ₃ (7.4 g, 54 mmol), palladium acetate (0.31 g, 1.4 mmol) and 1,3-bis (diphenylphosphine) propane (0.75 g, 1 mmol). , 82 mmol). The mixture was placed in an autoclave under CO (0.8 MPa) and heated at 50 ° C. After 4 hours, the mixture was poured into water (10 volumes) and extracted four times with ethyl acetate. The organic phase was washed with aqueous NaCl and decolourised with charcoal, filtered and dried to give a solid which was purified by flash chromatography (eluent: petrolatum / ethyl acetate 1: 1) to give 5 g of the title compound (yield: 49%).

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 8.22-7.55 (m, 3H); 4.91 (s, 2 H); 4.02 and 4.01 (2s, 6H).

- 79 · .. .. .. 79 79 ♦ 79 79 79 79 79,

Example 103 [4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-phthalazin-1-yl] -methanol

To a suspension of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxyphthalazine-1-carboxylic acid methyl ester (3.9 g, 103 mmol), prepared as described in Example 102, in DMF / CH ₃ OH (30/50 mL) under N ₂ was added portionwise NaBH ₄ (1.17 g, 30.9 mmol) over the course of

Stirring was continued for 2.5 hours. The mixture was cooled on ice and concentrated HCl was added to pH <1, then the mixture was dried and extracted three times with CH ₂ Cl ₂ , the organic phase was dewatered and dried to give 3.5. g of the title compound (yield: 97%). Melting point: 190-195 ° C.

¹ H-NMR (CDCl 3) δ: 8.51 (s, 2H); 8.20-7.10 (m, 3H); 5.22 (s, 2H, J = 4.4Hz); 4.90 (s, 2 H); 4.50 (t, IH); 4.01 (s, 3H).

Example 104

1- (3,5-Dichloropyridin-4-ylmethyl) -6-methoxy-4-morpholin-4-ylmethyl-phthalazine (Compound 56)

To a suspension of N ₂ [4- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazin-1-yl] -methanol (1.25 g, 3.6 mmol), prepared as described in Example 103 , in CH ₂ Cl ₂ (30 mL), triethylamine (0.75 mL, 5.4 mmol) was added. The mixture was cooled to 0-4 ° C and methanesulfonyl chloride (0.33 mL, 4.32 mmol) in CH ₂ Cl ₂ (5 mL) was added dropwise. After the addition was complete, the resulting solution was stirred for 1 hour at room temperature, cooled and morpholine (0.94 mL, 10.8 mmol) was added. The mixture was stirred at room temperature for 3 hours, then poured into 5% HCl. The acid phase was basified and extracted twice with CH ₂ Cl ₂ , then decolourised with charcoal, filtered through celite and dried to give a foam which was purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH 3 OH 98: 2). The resulting foam was triturated in ethyl ether to give 1.1 g of the title compound (yield: 52%). Melting point: 202.6 - 205.6 C. ¹ H-NMR (CDCl 3) δ: 8.49 (s, 2H); 8.14-7.52 (m, 3H); 4.87 (s, 2 H); 4.10 (s. 2H); 4.00 (s. 3H); 3.68-3.64 (m, 4H); 2.57-2.52 (m, 4H).

Example 105

1- (3,5-Dichloropyridine-4-ylmethyl) -6-methoxy-4-pyrrolidin-1-ylmethyl-phthalazine (Compound 57)

Using essentially the same procedure as described in Example 104, starting from [4- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazin-1-yl] methanol (1.75 g, 5 mmol), produced as described in Example 103, in CH ₂ Cl ₂ (30 mL), triethylamine (1 mL,

7.5 mmol), methanesulfonyl chloride (0.47 mL, 6 mmol) in CH ₂ Cl ₂ (5 mL), and pyrrolidine (1.24 mL, 15 mmol) gave 1.04 g of the title compound (yield: 51%). Melting point: 174.6-176.6 ° C.

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 8.12-7.49 (m, 3H); 4.86 (s, 2 H); 4.19 (s, 2 H); 4.00 (s. 3H); 2.63-2.57 (m, 4H); 1.77-1.71 (m, 4H).

Example 106

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4- (3-morpholin-4-yl-prop-1-invl) -phthalazine (Compound 58)

To a solution of N ₂ 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (1.3 g, 3.7 mmol), prepared as described in Example 45, in dry DMF ( 50 mL), K ₂ CO ₃ (1.28 g, 9.25 mmol), palladium acetate (0.017 g, 0.074 mmol), CuI (0.028 g, 0.15 mmol) and triphenylphosphine (0.58 g, 0.22 mmol). After stirring for 30 min, 4-prop-2-ynyl-morpholine (1.16 g, 9.25 mmol) was added and the mixture was stirred overnight, poured into water (10 volumes) and extracted twice with ethyl acetate. After decolourising with charcoal, the mixture was filtered through

- 81 celite, concentrated and triturated in ethyl ether; 1.15 g of the title compound was obtained (yield: 70%). Melting point: 171.7 172.7 ° C (dec.).

¹ H-NMR (CDCl 3) δ: 8.49 (s, 2H); 8.13-7.54 (m, 3H); 4.88 (s, 2 H); 4.02 (s, 3H); 3.77-3.72 (m, 4H); 3.72 (s, 2 H); 2.76-2.71 (m, 4H).

Example 107

4-14- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-phthalazin-1-yl-2-methyl-but-3-yn-2-ol (Compound 59)

Using essentially the same procedure described in Example 106, starting from 4-chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (1 g, 2.8 mmol), prepared as described above. in Example 45, in dry DMF (25 mL), K ₂ CO ₃ (0.97 g, 7 mmol), palladium acetate (0.013 g, 0.056 mmol), CuI (0.021 g, 0.11 mmol), triphenylphosphine (0.044) g, 0.17 mmol) and 2-methyl-3-butin-2-ol (0.59 g, 7 mmol) gave 0.93 g of the title compound. Melting point: 203.6 - 205.6 ° C (dec.).

¹ H-NMR (CDCl 3) δ: 8.48 (s, 2H); 8.10-7.52 (m, 3H); 4.87 (s, 2 H); 4.00 (s. 3H); 2.65 (s, 1 H); 1.70 (s, 6H).

Example 108

1- (3,5-Dichloropyridin-4-ylmethyl) -4-ethynyl-6-methoxyphthalazine (Compound 60)

In a Claisen flask under N ₂ , to a suspension of 4- [4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxyphthalazin-1-yl] -2-methyl-but-3-yn-2-ol (0 (4 g, 1 mmol), prepared as described in Example 107, in dry toluene (50 mL), a catalytic amount of NaH was added, the mixture was refluxed, then distilled to half volume. The mixture was allowed to stand overnight, then poured into water. *

Extracted three times with CH 2 Cl 2, decolourised with charcoal, filtered through celite and dried to give a solid which was triturated in ethyl ether to give 0.21 g of the title compound. Melting point: 210.3-211.3 ° C (dec.).

¹ H-NMR (CDCl 3) δ: 8.49 (s, 2H); 8.14-7.54 (m, 3H); 4.90 (s, 2 H); 4.03 (s, 3H); 3.62 (s, 1 H).

Example 109

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-phthalazine-1-carboxylic acid amide (Compound 61)

Excess NH ₃ was bubbled into a suspension of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxyphthalazine-1-carboxylic acid methyl ester (1.6 g, 4.23 mmol), prepared as described in Example 102, in dry CH ₃ OH, with stirring under dry N ₂ at 10 ° C, and the mixture was left at room temperature for 3 hours, then refluxed for 1.5 hours in an NH ₃ saturated environment. After 2 days at room temperature, the mixture was dried to give 1.48 g of the title compound (yield: 67.7%).

¹ H-NMR (DCDIs) δ: 9.00-7.57 (m, 3H); 8.53 (s, 2 H); 4.95 (s, 2 H); 4.05 (s, 3H).

Example 110

4- (3,5-Dichloro-pyridin-4-ylmethyl) -7-methoxy-phthalazine-1-carbonitrile (compound 62)

A solution of 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxyphthalazine-1-carboxylic acid amide (1.06 g, 2.92 mmol), prepared as described in Example 109, in pyridine (80 mL). and trifluoromethanesulfonic anhydride (1.65 mL, 11.67 mmol), under dry N ₂ , at room temperature, was stirred for 3 hours, then dried, transferred to

9 9

9

9 9

9

9

- 82 9 «• 9 9

9

9999

9

9999 8 9

CH ₂ Cl ₂ , washed with citric acid, 5% NaOH and water. The organic phase was drained and dried to give a solid which was purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH 3 OH 99: 1) to give 0.33 g of the title compound (yield: 33%).

¹ H-NMR (CDCl 3) δ: 8.52 (s, 2H); 8.22 (d, 1H, J = 9.2Hz); 7.69 (dd, IH); 7.47 (d, 1H, J = 2.5Hz); 4.96 (s, 2 H); 4.07 (s, 3H).

Example 111

4- _(3-l 5 Dichlorpvridin-4-ylmethyl-7-methoxy-1-phenyl-1,2-dihvdroftalazin (Compound 63)

To 1- (3,5-dichloropyridin-4-ylmethyl) -4-phenyl-6-methoxyphthalazine (0.5 g, 1.26 mmol), prepared as described in Example 47, in THF (40 mL) was added. platinum oxide hydrate (catalytic amount) was added, and the mixture was hydrogenated in a Parr apparatus at 0.4 MPa with stirring for 4 days and at 50 ° C for 24 hours. The catalyst was filtered off and the mixture was dried to give a solid which was purified by flash chromatography (eluent: petrolatum / ethyl acetate 8: 2) to give 0.16 g of the title compound (yield: 32%). Melting point: 175.3 - 177.3 ° C.

¹ H-NMR (CDCl 3) δ: 8.45 (s, 2H); 7.42-6.27 (m, 8H); 5.78 (s, 1 H); 5.21 (s, 1 H); 4.36-4.15 (m, 2 H); 3.71 (m, 3H).

Example 112

4- (3,5-Dichloro-pyridin-4-ylmethyl) -2-methanesulfonyl-7-methoxy-1-phenyl-1,2-dihydrophthalazine (Compound 64)

To 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1-phenyl-1,2-dihydrophthalazine (0.16 g, 0.4 mmol), prepared as described in Example 111, in dry CH ₂ Cl ₂ (10 mL), under N ₂ , was added triethylamine (0.1 mL, 0.72 mmol) and methanesulfonyl chloride (0.037 mL, _o .).

- 84 - ·· ········ · ·

0.48 mmol). The mixture was stirred at room temperature overnight, additional triethylamine (0.05 mL, 0.36 mmol) and methanesulfonyl chloride (0.02 mL, 0.24 mmol) were added. After 4 hours, methanesulfonyl chloride (0.04 mL, 0.48 mmol) and 1,8-diazabicyclo [5.4.0] undec-7-ene (0.085 mL, 0.56 mmol) were added. After 24 h, it was poured into water and the aqueous phase was separated and extracted with CH 2 Cl 2. The organic phase was washed with dilute citric acid, dehydrated and dried to give a solid which was purified by flash chromatography (eluent: petrolatum / ethyl acetate 7: 3). The solid obtained was triturated in ethyl ether to give 0.21 g of the title compound (quantitative yield). Melting point: 222.6 - 224.4 ° C.

¹ H-NMR (CDCl 3) δ: 8.47 (s, 2H); 7.60 (d, 1H, J = 8.7Hz); 7,287.24 (m, 5H); 6.94 (dd, IH); 6.64 (d, 1H, J = 2.6Hz); 6.23 (s, 1 H); AB system; Va = 4.51, Vb = 4.33, Jab = 16.3 Hz); 3.80 (s, 3H); 2.21 (s, 3H).

Example 113

4- (6-Methoxy-4-oxo-3,4-dihydrophthalazin-1-yl) butyric acid methyl ester (Compound 65)

To a solution of 3-formylpropionic acid methyl ester (13.4 g, 0.1155 mol) and (5-methoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenylphosphonium bromide (55.6 g, 0.11 mol) ), prepared as described in Example 36, in CH ₂ Cl ₂ (440 mL) in a water bath, triethylamine (16.1 mL, 0.1155 mol) was added dropwise. After the addition was complete, the mixture was stirred at room temperature overnight, then washed with water, dewatered and dried to give a residue which was taken up in ethanol (180 mL). The solution was cooled and hydrazine monohydrate (11 g, 0.22 mol) was added. The precipitate formed was filtered. After 6 days, the mother liquors gave a precipitate which, combined with the preceding material, gave 23 g of the title compound (yield: 75.5%). Melting point: 175-177 ° C.

• · ·· ··

@ ¹ H-NMR (DMSO) .delta .: 12.40 (broad, 1H); 7.96-7.45 (m, 3H); 3.92 (s, 3H); 3.57 (s. 3H); 2.92-2.85 (m, 2H); 2.48-2.40 (m, 2 H); 2.00-1.84 (m, 2 H).

Example 114

4- (4-Chloro-6-methoxy-phthalazin-1-yl) -butyric acid methyl ester

A suspension of 4- (6-methoxy-4-oxo-3,4-dihydrophthalazin-1-yl) -butyric acid methyl ester (9.23 g, 0.034 mol), prepared as described in Example 113, and POCl 3 (1.87) ml, 0.2 mol) in acetonitrile (100 ml) was refluxed. After 1.5 h, the mixture was dried, taken up in water and neutralized with NaHCO 3. The mixture was extracted with ethyl acetate and the organic phase was dried to give a residue which was purified by flash chromatography (eluent: petrolatum / ethyl acetate 7: 3). 7.4 g of the title compound were obtained (yield: 63.9%).

Example 115

4- (6-Methoxy-4-thiazol-2-yl-phthalazin-1-yl) -butyric acid methyl ester (Compound 66)

Using essentially the same procedure described in Example 48, starting from 4- (4-chloro-6-methoxy-phthalazin-1-yl) -butyrate (12.5 g, 0.0424 mol), prepared as described above. in Example 114, zinc (17.3 g), 2-bromothiazole (24.3 g, 0.1484 mol), palladium acetate (0.476 g, 2.12 mmol), triphenylphosphine (1.67 g, 6.36 mmol) ), 3.8 g of the title compound was obtained (yield: 12.6%). M.p .: 136-137 ° C.

¹ H-NMR (CDCl 3) δ: 9.24 (d, 1H, J = 2.7); 8.11 (d, 1H, J = 9.1); 8.04 (d, 1H, J = 3.3); 7.53-7.47 (m, 2H); 4.03 (s, 3H); 3.67 (s, 3H); 3.42-3.34 (m, 2 H); 2.57-2.50 (m, 2 H); 2.33-2.19 (m, 2 H).

4 4 4

44443 4 4

- 86 Example 116

N-Hydroxy-4- (6-methoxy-4-thiazol-2-yl-phthalazin-1-yl) -butyramide (Compound 67)

To a solution of sodium (388 mg, 16.9 mmol) in CH ₃ OH (6 mL) was added hydroxylamine hydrochloride (1.3 g, 18.71 mmol). The resulting solution was added to a solution of 4- (6-methoxy-4-thiazol-2-yl-phthalazin-1-yl) -butyric acid methyl ester (1.5 g, 4.4 mmol), prepared as described in Example 115, in CH 3 OH (30 mL), and the mixture was stirred at room temperature for 2 days, then under reflux for 7 h. NH 2 OH.HCl (1.6 g) and sodium (450 mg) were added and the mixture was refluxed. After one night, additional NH ₂ OH.HCl (0.9 g) and sodium (257 mg) were added and the mixture was refluxed for 24 hours. The precipitate was filtered off and the mother liquors were dried, taken up in water and filtered. The obtained solid was purified by chromatography (eluent: CH 2 Cl 2 / CH 3 OH 95: 5) to give a solid which was suspended in water, dissolved in 10% NaOH and precipitated with saturated ammonium chloride solution to give 430 mg of the title compound (yield: 28). , 6%). Melting point: 189-190 ° C (dec.).

¹ H-NMR (DMSO) δ: 10.36 and 8.72 (2 broad, 2H); 9.15 (d, 1H, J = 2.6); 8.32 (d, 1H, J = 9.2); 8.19 (d, 1H, J = 3.3); 7.99 (d, IH); 7.70 (dd, IH); 3.99 (s, 3H); 3.35-3.27 (m, 2 H); 2.18-1.97 (m, 4H).

Example 117

4- (6-Methoxy-4-thiazol-2-yl-phthalazin-1-yl) -butyramide (Compound 68)

To a suspension of 4- (6-methoxy-4-thiazol-2-yl-phthalazin-1-yl) -butyric acid methyl ester (1.2 g, 3.5 mmol), prepared as described in Example 115, in NH 3 / THF (10 mL) was added THF (15 mL), the mixture was placed in an autoclave at 85 ° C. After one night the mixture was dried,

Transferred to CH ₃ OH (30 mL) and saturated with NH ₃ at 0 ° C, then heated to 85 ° C for 44 h. The mixture was cooled and dried to give a residue which was purified by flash chromatography (eluent: CH 2 Cl 2 / CH 3 OH 95). : 5 to 90: 10). The solid was triturated in CH ₂ Cl ₂ to give 840 mg of the title compound (yield: 73.1%).

¹ H-NMR (DMSO) δ: 9.15 (d, 1H, J = 2.6); 8.35 (d, 1H, J = 9.2); 8.19 (d, 1H, J = 3.2); 7.99 (d, IH); 7.69 (dd, IH); 7.30 and 6.77 (2s broad, 2H); 3.99 (s, 3H); 3.35-3.28 (m, 2 H); 2.26-2.19 (m, 2 H); 2.09-1.95 (m, 2 H).

Example 118

4- (6-Methoxy-4-thiazol-2-yl-phthalazin-1-yl) -butyric acid (compound

69)

4- (6-Methoxy-4-thiazol-2-yl-phthalazin-1-yl) -butyric acid methyl ester (1.2 g, 3.5 mmol), prepared as described in Example 115, was dissolved in hot CH ₃ OH (30 mL) and 10% NaOH (2.8 mL, 7 mmol) was added, and the mixture was refluxed. After 30 min, the mixture was cooled, dried, taken up in water and extracted with CH ₂ Cl ₂ . The aqueous phase was decolourised with activated carbon and acidified with acetic acid. The precipitate was filtered, washed with water and dried, yielding 0.95 g of the title compound (yield: 82.4%). Melting point: 218-220 ° C.

¹ H-NMR (DMSO) δ: 12.11 (s, 1H); 9.13 (d, 1H, J = 2.7); 8.32 (d, 1H, J = 9.2); 8.18 (d, 1H, J = 3.3); 7.98 (d, IH); 7.67 (dd, IH); 3.98 (s, 3H); 3.37-3.29 (m, 2 H); 2.45-2.38 (m, 2 H); 2.11-1.96 (m, 2 H).

• 9 9 99 9999999 9

88 Example 119

3- (6-Methoxy-4-oxo-3,4-dihydrophthalazin-1-yl) -propionic acid ethyl ester (Compound 70)

Using essentially the same procedure as described in Example 113, starting from potassium 2-ethoxycarbonylethenolate (771 mg, 5 mmol), (5-methoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) triphenylphosphonium bromide (2, 2-ethoxycarbonylethenolate). 53 g, 5 mmol), prepared as described in Example 36, CH 2 Cl 2 (20 mL) and hydrazine monohydrate (250 mg, 5 mmol) in ethanol (20 mL) gave 0.52 g of the title compound.

¹ H-NMR (CDCl 3) δ: 7.80-7.35 (m, 3H); 4.13 (q, 2 H); 3.95 (s, 3H); 3.28-2.77 (m, 4H); 1.23 (t, 3H, J = 7.1).

Example 120

3- (3-Methanesulfonyl-6-methoxy-4-oxo-3,4-dihydro-phthalazin-1-yl) -propionic acid ethyl ester (Compound 71)

A suspension of 3- (6-methoxy-4-oxo-3,4-dihydro-phthalazin-1-yl) -propionic acid ethyl ester (4 g, 0.0 145 mol), prepared as described in Example 119, and NaH ( 0.638 g, 0.016 mol) in acetonitrile (80 mL) was heated to 55 ° C. After 2 h, the mixture was cooled, methanesulfonyl chloride (1.35 mL, 0.017 mol) was added, and the mixture was stirred at room temperature overnight. K 2 CO 3 (2 g, 0.0145 mol) and methanesulfonyl chloride (1.12 mL, 0.0145 mol) were added and the mixture was stirred overnight, then dried and partitioned between water and CHCl 3. The organic phase was washed with K 2 CO 3 solution, dewatered and dried to give a residue which was purified by flash chromatography (eluent: CH ₂ Cl / ethyl acetate 95 5). The solid obtained was crystallized from acetonitrile (5 ml) to give 0.9 g of the title compound (yield: 17.5%).

¹ H-NMR (DMSO) δ: 7.81 (d, 1H); 7.78 (d, IH); 4.41 (dd, 1 H,

89 89 J = 8.9 Hz, 2.8 Hz); 4.15 (q, 2 H); 3.95 (s, 3H); 3.55 (s, 3H); 3.31-3.24 (m, 2 H); 2.90-2.83 (m, 2 H); 1.26 (t, 3H, J = 7.1).

Example 121

3- (4-Chloro-6-methoxy-phthalazin-1-yl) -propionic acid ethyl ester

Using essentially the same procedure as described in Example 114, starting from 3- (6-methoxy-4-oxo-3,4-dihydrophthalazin-1-yl) -propionic acid ethyl ester (552 mg, 2 mmol), prepared as as described in Example 119, and POCl ₃ (1.12 mL, 12 mmol) in acetonitrile (5 mL) gave 0.5 g of the title compound (yield: 84.8%).

Example 122

3- (6-Methoxy-4-thiazol-2-yl-phthalazin-1-yl) -propionic acid ethyl ester (Compound 72)

Using essentially the same procedure described in Example 48, starting from 3- (4-chloro-6-methoxy-phthalazin-1-yl) -propionic acid ethyl ester (4.5 g, 0.0153 mol), prepared as was described in Example 121, zinc (2.2 g), 2-bromothiazole (5 g, 0.0305 mol), palladium acetate (180 mg, 0.8 mmol), triphenylphosphine (630 mg, 2.4 mmol), 1.4 g of the title compound was obtained (yield: 21%). Melting point: 153-154 ° C.

¹ H-NMR (CDCl ₃ ) δ: 9.26 (d, 1H, J = 2.7); 8.11 (d, 1H, J = 9.2); 8.09 (d, 1H, J = 3.2); 7.52 (dd, IH); 7.50 (d, IH); 4.15 (q, 2 H); 4.04 (s, 3H); 3.68-3.61 (m, 2 H); 3.16-3.09 (m, 2 H); 1.24 (t, 3H, J = 7.1).

• · • ·

- 90 Example 123

3- (6-Methoxy-4-thiazol-2-yl-phthalazin-1-yl) -propionic acid (Compound 73)

Using essentially the same procedure as described in Example 118, starting from 3- (6-methoxy-4-thiazol-2-yl-phthalazin-1-yl) -propionic acid ethyl ester (1.05 g, 3.06 mmol) ), prepared as described in Example 122, CH ₃ OH (25 mL) and 10% NaOH (2.5 mL, 6.25 mmol) yielded 0.8 g of the title compound (yield: 82.9%) . Melting point: 228-229 ° C.

¹ H-NMR (DMSO) δ: 12.7 (s, 1H); 9.14 (d, 1H, J = 2.6); 8.33 (d, 1H, J = 9.3); 8.19 and 7.99 (2d, 2H, J = 3.3); 7.69 (dd, IH); 3.99 (s, 3H); 3.60-3.54 (m, 2 H); 2.96-2.89 (m, 2 H).

Example 124

N -hydroxy-3- (6-methoxy-4-thiazol-2-yl-phthalazin-1-yl) -propionamide (Compound 74)

A solution of 3- (6-methoxy-4-thiazol-2-yl-phthalazin-1-yl) propionate (200 mg), prepared as described in Example 123, H2SO4 (3 drops) and _CH3 OH (15 ml ) was refluxed overnight, decolourised with activated carbon, neutralized with Na ₂ CO ₃ and dried. The residue was partitioned between ethyl acetate and water, the organic phase was dewatered and dried to give residue (A). In another flask, to sodium (47 mg, 2.04 mmol) in CH ₃ OH (2 mL) was added NH ₂ OH.HCl (142 mg, 2.04 mmol) and residue (A) in CH ₃ OH (12 mL) ). The mixture was refluxed under N _{2 for} 2 hours, then stirred at room temperature for 4 days. A precipitate formed, the mixture was cooled in an ice-water bath, filtered, washed with water and dried, yielding 75 mg of the title compound (yield: 53.4%).

¹ H-NMR (DMSO) δ: 10.51 and 8.72 (2s, 2H); 9.16 (d, 1H, J = 2.6); 8.33 (d, 1H, J = 9.1); 8.20 (d, 1H, J = 3.3); 7.99 (d, IH); 7.71 (dd, IH);

4.00 (s. 3H); 3.57 (t, 2H, J = 7.3); 2.64 (t, 2 H).

Example 125

Sodium 1- [4- (3 ₁ 5 ', 4-chloro-pyridin-4-ylmethyl) -7-methoxy-phthalazin-1-yl] -pyrrolidine-2-carboxylate (Compound 75)

To a suspension of L (-) proline (5.2 g, 0.045 mol) in acetonitrile (50 mL) under N 2 was added NaH (0.9 g, 0.0375 mol) and the mixture was stirred at room temperature for 1 hour. 4-Chloro-1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine (5.32 g, 0.015 mol), prepared as described in Example 45, was added, and the mixture was refluxed. After 2 days the mixture was dried under vacuum, the residue was taken up in water and extracted with CH 2 Cl 2. The alkaline aqueous phase was decolourised with activated carbon, filtered through celite, and the filtrate was acidified (pH = 4) with 10% HCl and extracted three times with CH 2 Cl 2. The organic phase was dehydrated over Na 2 SO ₄ and dried. The residue was converted into the sodium methoxide salt, dissolved in CH 3 OH and dried. The salt was treated with hot tert-butyl methyl ether (100 mL), cooled in water / ice and filtered to give 6.4 g of the title compound (yield: 84.3%).

¹ H-NMR (DMSO) δ: 8.10 (s, 2H); 8.10 (d, 1H, J = 9.0); 7.79 (d, 1H, J = 2.4); 7.45 (dd, IH); 4.64 (s, 2 H); 4.65-4.58 (m, IH); 3.92 (s, 3H); 3.78-3.66 (m, 2 H); 2.24-1.66 (m, 4H).

Example 126

4-Furan-2-ylmethyl-7-methoxy-2H-phthalazin-1-one (compound 76)

To a mixture of (5-methoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenylphosphonium bromide (8 g, 15.83 mmol), prepared as described in Example 36, CH 2 Cl 2 (100 mL) and furfural (1.31 mL, 15.83 mmol), ice-cooled under N ₂ , triethylamine (2.2 mL, 15.83 mmol) was added dropwise. Cooling was discontinued and the mixture was stirred for 1.5 hours, then washed with water, dehydrated over Na 2 SO ₄

- 92 • 4 * 4 4 4 9 4 449 4 44 9 49 9444944 4 9 9 49 »4» 4 4444 »4« and dried. The residue was dissolved in CH ₃ OH (50 mL) and hydrazine monohydrate (2.3 mL, 47.49 mmol) was added. The mixture was refluxed for 1 hour, then concentrated to half volume. The precipitate crystallized and was dried under vacuum at 40 ° C to give 1.82 g of the title compound (yield: 45%).

¹ H-NMR (DMSO) δ: 12.50 (s, 1H); 7.96-7.43 (m, 3H); 7.51-6.20 (m. 3H); 4.27 (s, 2 H); 3.91 (s, 3H).

Example 127

4-Chloro-1-furan-2-ylmethyl-6-methoxy-phthalazine

To a suspension, under N ₂ , 4-furan-2-ylmethyl-7-methoxy-2H-phthalazin-1-one (1.8 g, 7.02 mmol), prepared as described in Example 126, in acetonitrile (40 mL). mL), POCl ₃ (3.27 mL, 35.1 mmol) was added with stirring, and the mixture was refluxed for 1 hour, then dried. The residue was suspended in water (50 mL), NaHCO ₃ was added until alkalinity and the mixture was stirred for 30 min, and then extracted with CH ₂ Cl ₂ . The organic phase was drained over Na ₂ SO ₄ and dried to give a residue which was mixed with petrolatum and filtered. The obtained solid was dried under vacuum at 40 ° C to give 1.72 g of the title compound (yield: 89%).

Example 128

1-Furan-2-ylmethyl-6-methoxy-4-phenyl-phthalazine (Compound 77)

To a solution of N ₂ ZnCl ₂ in THF 0.5M (18.35 mL, 9.18 mmol) was added dropwise 2M phenyllithium (4.37 mL, 8.74 mmol) with stirring at 0 ° C. The mixture was left at room temperature for 1 hour, then 4-chloro-1-furan-2-ylmethyl-6-methoxyphthalazine (1.2 g, 4.37 mmol), prepared as described in Example 127, palladium acetate was gradually added. (49 mg, 0.218 mmol) and triphenylphosphine (114.4 mg, 4.36 mmol). The mixture was refluxed for 45 min, then diluted.

- 93 - · ethyl acetate and washed with water. The organic phase was dehydrated over Na ₂ SO ₄ and dried. The residue was purified by flash chromatography (eluent: petrolatum / ethyl acetate 1: 1). The obtained oil crystallized from ethyl ether (15 mL) to give 480 mg of the title compound (yield: 34%). Melting point: 128 - 129 ° C.

¹ H-NMR (CDCl 3) δ: 8.14 (d, 1H, J = 9.2 Hz); 7.76-7.50 (m, 5H); 7.44 (dd, 1H, J = 2.6 Hz); 7.31-7.29 (m, 2 H); 6.28-6.25 (m, IH); 6.10 (d, 1H, J = 3.3Hz); 4.72 (s, 2 H); 3.82 (s, 3H).

Example 129

-Furan-2-ylmethyl-6-methoxy-4-pyrrolidin-1-yl-phthalazine (Compound 78)

To a solution of 4-chloro-1-furan-2-ylmethyl-6-methoxyphthalazine (500 mg, 1.82 mmol), prepared as described in Example 127, in DMF (10 mL) was added pyrrolidine (0, 91 mL, 10.92 mmol), and the mixture was heated to 60 ° C overnight. The mixture was diluted with ethyl acetate, washed three times with water, dried over Na ₂ SO ₄ and dried. The residue was crystallized from ethyl ether / petrolatum 1: 1 (10 mL) to give 420 mg of the title compound (yield: 75%). M.p .: 148-149 ° C.

¹ H-NMR (CDCl 3) δ: 7.91 (d, 1H, J = 9.1 Hz); 7.48 (d, 1H, J = 2.6Hz); 7.34-7.26 (m, 2 H); 6.24-6.21 (m, IH); 6.01 (d, 1H, J = 3.3Hz); 4.49 (s, 2 H); 3.91 (s, 3H); 3.87-3.81 (m, 4H); 2.03-1.96 (m, 4H).

Example 130

7-Methoxy-4-pyridin-4-ylmethyl-2H-phthalazin-1-one (Compound 79)

To a solution of (5-methoxy-3-oxo-1,3-dihydroisobenzofuran-1-yl) -triphenylphosphonium bromide (43.5 g, 0.0861 mol), prepared as described in Example 36, and 4-pyridinecarboxaldehyde ( 9.22 g, 0.0861 mole) of CH ₂ Cl ₂ (300 mL) was added dropwise triethylamine · · * 300 300 iet iet 300 300 iet ·

9 9 9 9 9 9 9 9 9

94 (12 ml, 0.0861 mol), maintaining the temperature in the water bath. After stirring overnight at room temperature, the mixture was washed with water, decolourised, dewatered and dried. The residue was suspended in CH ₃ OH (135 mL) and hydrazine monohydrate (12.55 mL, 0.2583 mol) was added. The mixture was refluxed for 1 hour, then cooled in water / ice, filtered and dried. The residue was crystallized in CH 3 OH to give 8 g of the title compound. The mother liquors were dried and the residue was triturated in CH ₃ OH (50 mL) to give an additional 3.5 g of the title compound (total yield: 50%) ¹ H-NMR (DMSO) δ: 12.54 (s, 1H) ; 8.46-7.27 (m, 7H); 4.28 (s, 2 H); 3.90 (s, 3H).

Example 131

4-Chloro-6-methoxy-1-pyridin-4-ylmethyl-phthalazine

A suspension of 7-methoxy-4-pyridin-4-ylmethyl-2H-phthalazin-1-one (11.4 g, 42.65 mmol), prepared as described in Example 130, and POCl 3 (7.95 ml, 85, 30 mmol) in acetonitrile (110 mL) was refluxed for 1.5 h additional POCl 3 (40 mL) was added and refluxed overnight. The mixture was dried, dissolved in water, neutralized with NaHCO 3, extracted with CH ₂ Cl ₂ and dried. The residue was purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH ₃ OH 97: 3) to give 2.9 g of the title compound (yield: 23.8%).

¹ H-NMR (CDCl 3) δ: 8.48-7.15 (m, 4H); 7.85-7.40 (m, 3H); 4.62 (s, 2 H); 4.00 (s, 3H).

· · * * * * * * · Φ φ φ φ <φ <·

- 95 ··

Example 132

6-Methoxy-1-pyridin-4-ylmethyl-4-pyrrolidin-1-yl-phthalazine (Compound 80)

Using essentially the same procedure as described in Example 129 but under N 2, starting from 4-chloro-6-methoxy-1-pyridin-4-ylmethyl-phthalazine (200 mg, 0.7 mmol), prepared as described in of Example 131, in DMF (5 mL) and pyrrolidine (0.17 mL, 2.1 mmol) gave 180 mg of the title compound (yield: 65%).

¹ H-NMR (DMSO) δ: 8.90-8.86 (m, 2H); 8.27 (d, 1H, J = 9.1Hz); 8.00-7.95 (m, 3H); 7.78 (dd, 1H, J = 2.1Hz); 4.88 (s, 2 H); 4.05-3.96 (m, 4H); 4.02 (s, 3H); 2.09-2.02 (m, 4H).

Example 133

3- [2- (2-Methoxyethoxy) -ethoxy] propyne

To a solution, under N ₂ , 2- (2-methoxyethoxy) -ethanol (4.9 mL, 41.61 mmol) in dry THF (50 mL), was added portionwise, at room temperature, with stirring 60% NaH in oil (1.66 g, 41.61 mmol). The mixture was stirred for 30 min, then added dropwise to a solution of 80% 3-brompropine in toluene (5.1 mL, 45.77 mmol), and stirred for 1 h, then dissolved in ethyl ether, washed three times with water, dehydrated over Na 2 SO 4 and dried to give 5 g of the title compound as an oil (yield: 76%).

¹ H-NMR (CDCl 3) δ: 4.18 (d, 2HJ = 2.4 Hz); 3.71-3.51 (m, 8H); 3.36 (s, 3H); 2.40 (t, 1 H).

Example 134

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-5- {3- [2- (2-methoxy-ethoxy) -ethoxy-prop-1-ynyl] -phthalazine (Compound 81)

To a suspension, under N 2, 3- [2- (2-methoxyethoxy) -ethoxy] -propine (810 mg, 5.12 mmol), prepared as described in Example 133, was

- 99 99 99 99 99 99 99 999 99

9 9 9 9999 * 9 9

9,999,999 trifluoromethanesulfonic acid 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazin-5-yl ester (2 g, 4.27 mmol), prepared as described in Example 73, and diethylamine ( 40 mL), with stirring, bis (triphenylphosphine) PdCl ₂ (60 mg, 0.085 mmol) and CuI (16.18 mg, 0.085 mmol) The mixture was refluxed for 6 h then dried. The residue was purified by flash chromatography (eluent: ethyl acetate) to give a solid which was taken up in ethyl ether, filtered and dried under vacuum at 40 ° C to give 940 mg of the title compound (yield: 46%).

¹ H-NMR (DMSO) δ: 9.62 (s, 1H); 8.68 (s, 2 H); 8.59-7.99 (2d, 2H, J = 9.3 Hz); 4.99 (s, 2 H); 4.59 (s, 2 H); 4.09 (s, 3H); 3.74-3.40 (m, 8H); 3.21 (s, 3H).

Example 135

Benzyl-3- [1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxyphthalazine-5-1H-allyl-methylamine (Compound 82)

Benzyl- {3- [1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl] -prop-2-ynyl} -methylamine solution (0.17 g, 0.36 mmol) ), prepared as described in Example 74, in THF (5 mL) and 10% Pd / C (0.07 g) was placed in a Parr apparatus at 50 psi for 1.5 h. The mixture was filtered through celite and dried to give an oil which was purified by flash chromatography (eluent: CH ₂ Cl ₂ / CH ₃ OH / NH ₃ 98: 2: 0.2, then 98: 2: 0.5). The obtained oil crystallized from isopropyl ether to give 0.105 g of the title compound (yield: 61%).

¹ H-NMR (CDCl ₃ ) δ: 9.40 (s, 1H); 8.51 (s, 2 H); 8.17 and 7.62 (2d, 2H, J = 9.2 Hz); 7.23-7.11 (m, 5H); 6.65 (dt, IH); 6.29 (dt, 1H, J = 11.5 Hz, J = 6.6 Hz); 4.91 (s, 2 H); 3.98 (s, 3H); 3.34 (s, H); 2.83 (dd, 2H, J ₂ H = 1.65 Hz).

• ♦

Example 136

Methanesulfonic acid prop-2-yl ester

A solution of prop-2-in-1-ol (5 g, 89.19 mmol) in CH ₂ Cl ₂ (25 mL) was cooled to 0-5 ° C and triethylamine (13.6 mL, 98.11 mmol) was added. ) and a solution of methanesulfonyl chloride (7.6 mL, 98.11 mmol) in CH ₂ Cl ₂ (20 mL). After 2 hours the mixture was placed in the refrigerator overnight. The mixture was washed with water and the organic phase was dried to give 6.88 g of the title compound.

¹ H-NMR (CDCl 3) δ: 4.82 (d, 2H, J = 2.4 Hz); 3.11 (s, 3H); 2.68 (t,

1H).

Example 137

4-Prop-2-ynylmorpholine

A mixture of propane-2-ynyl methanesulfonic acid ester (3.44 g, 25.7 mmol), prepared as described in Example 136, morpholine (5.4 g) and Na ₂ CO ₃ (3.3 g, 30.84). mmol) in ethanol was stirred at room temperature for 6 hours. The solid was filtered off and the solution was concentrated and poured into water. The mixture was acidified with HCl, extracted with ethyl ether, basified with NaHCO ₃ and extracted with ethyl acetate. The organic phase was dehydrated over Na ₂ SO ₄ and dried to give 3.87 g of the title compound, which was used as such in the next step.

Example 138

1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-5- (3-morpholin-4-yl-prop-1-ynyl) -phthalazine (Compound 83)

Using essentially the same procedure as described in Example 74, starting from trifluoromethanesulfonic acid 1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl ester (2 g, 4.27 mmol) ,

• φ

98, 4-prop-2-ynyl-morpholine (640 mg, 5.12 mmol), prepared as described in Example 137, diethylamine (-). 40 mL), bis (triphenylphosphine) PdCl ₂ (60 mg, 0.0854 mmol) and CuI (16 mg, 0.0854 mmol) gave 728 mg of the title compound (yield: 38.6%).

¹ H-NMR (CDCl 3) δ: 9.76 (s, 1H); 8.50 (s, 2 H); 8.16 and 7.60 (2d, 2H); 4.88 (s, 2 H); 4.08 (s, 3H); 3.80-3.75 (m, 4H); 3.72 (s, 2 H); 2.732.68 (m, 4H).

Example 139

6-Methoxy-5-trimethylsilanylethynyl-2H-phthalazin-1-one

A solution of trifluoromethanesulfonic acid 6-methoxy-1-oxo-1,2-dihydrophthalazin-5-yl ester (4.52 g, 13.94 mmol), prepared as described in Example 59, trimethylsilylethine (3.94 mL, 27 mL), 88 mmol), bis (triphenylphosphine) PdCl ₂ (245 mg, 0.384 mmol) and triethylamine (6.56 mL) in DMF (30 mL) were heated at 60 ° C for 4 h. Additional trimethylsilylethine (2 mL) was added and the mixture The mixture was heated at 90 ° C for 3 hours. After evaporation of the solvent, the residue was taken up in ethyl acetate and the solid was filtered off. After evaporation, the residue was taken up in diethyl carbonate, washed with water, dried again, and taken up in ethyl ether and filtered to give 2.87 g of the title compound (yield: 75.6%).

¹ H-NMR (DMSO) δ: 12.70 (s, 1H); 8.33 (s, 1 H); 8.22-7.58 (m, 2 H); 3.98 (s, 3H); 0.28 (s, 9H).

Example 140

5-Ethinyl-6-methoxy-2H-phthalazin-1-one

To a suspension of 6-methoxy-5-trimethylsilanylethynyl-2H-phthalazin-1-one (2.87 g, 10.20 mmol), prepared as described in Example 139, as described in Example 139. 999 999 • 9,999,999 • 9,999,999,999 9

999Π 999 999 99

- yy - 99 9999 99 9 99 9 in acetonitrile (50 ml), 32% NaOH (15 ml) was added and the mixture was stirred at room temperature for 1 hour. The mixture was neutralized with concentrated HCl and the precipitate was filtered off. The aqueous phase was filtered off with ethyl acetate, the organic phase was dried and taken up in acetone. The solid was filtered off and combined with the precipitate obtained previously to give 1.99 g of the title compound (yield: 97.3%).

¹ H-NMR (DMSO) δ: 12.68 (bs, 1H); 8.37 (s, 1 H); 8.24-7.60 (m, 2 H); 4.82 (s, 1 H); 4.00 (s, 3H).

Example 141

6-Methoxy-5-pyridin-4-ylethyl-2H-phthalazin-1-one

A suspension of 5-ethynyl-6-methoxy-2/7-phthalazin-1-one (1.9 g, 9.49 mmol), prepared as described in Example 140, 4-bromopyridine hydrochloride (2.214 g, 11.39 mmol) ), bis (triphenylphosphine) PdCl 2 (133 mg, 0.1998 mmol) and CuI (36 mg, 0.1898 mmol) in diethylamine (40 mL) was heated at 75 ° C for 1 h, cooled, diluted with water (40 mL) and the solid was filtered after stirring for 30 min. The solid was suspended in acetone, stirred for 15 min, filtered again and dried in an oven at 45 ° C under vacuum to give 2.263 g of the title compound (yield: 86%).

¹ H-NMR (DMSO) δ: 8.56 (s, 1H); 8.30-7.65 (m, 2 H); 4.05 (s, 3H).

Example 142

1-Chloro-6-methoxy-5-pyridin-4-ylethynyl-phthalazine

A suspension of N ₂ 6-methoxy-5-pyridin-4-ylethynyl-2H-phthalazin-1-one (2.26 g), prepared as described in Example 141, in POCl 3 (40 mL) was heated for 3 hrs. 85 ° C, then dried. The residue was treated in saturated NaHCO ₃ solution until alkaline

- 100 9 9

9 9

9 9

9999

9

9 9

9 9 9 • 9 9

9 9

9

PH, then filtered and extracted with ethyl acetate. The solid was dissolved in CH ₂ Cl ₂ / CH ₃ OH / ethyl acetate, dried over Na ₂ SO _4, and filtered. The solution was combined with the pre-obtained organic phase and dried to give a residue which was mixed with acetone and filtered to give 1.5 g of the title compound (yield: 62%).

¹ H-NMR (CDCl 3) δ: 9.87 (s, 1H); 8.70-7.70 (m, 4H); 8.41-8.04 (m, 2 H); 4.14 (s, 3H).

Example 143

1- _(3-l 5 Dichlorpvridin-4-ylmethyl) -6-methoxy-5-pyridin-4-vlethinvl-phthalazine (Compound 84)

Using essentially the same procedure described in Example 15, starting from 1-chloro-6-methoxy-5-pyridin-4-ylethynyl-phthalazine (1.38 g, 4.66 mmol), prepared as described in the Example. 142, 3,5-dichloro-4-methylpyridine (1.887 mg, 11.65 mmol), DMF (20 mL), NaH (466 mg, 11.65 mmol) gave 793 mg of the title compound.

¹ H-NMR (CDCl 3) δ: 9.83 (s, 1H); 8.68-8.64 (m, 2 H); 8.52 (s, 2 H); 8.25 and 7.66 (2d, 2H, J = 9.3 Hz); 7.51-7.48 (m, 2 H); 4.91 (s, 2 H); 4.15 (s, 3H).

Example 144

3- (3,5-Dichloro-pyridin-4-ylmethylene) -6-methoxy-1-thiazol-2-yl-1,3-dihydroisobenzofuran-1-ol

A milky solution of 2-bromothiazole (5.6 g, 34.2 mmol) in dry ethyl ether (17 mL) was added dropwise to a solution of n-butyllithium (14.4 mL, 36 mmol) in dry ethyl ether (50 mL), under N ₂ at -80 ° C. The reaction mixture was stirred at -80 ° C for 15 min, then a solution of 3- (3,5-dichloropyridin-4-ylmethylene) -6-methoxy-3H-isobenzofuran-1-one was added slowly. φ φ · · φ φ φ

101 (10 g, 31 mmol), prepared as described in Example 37, in dry THF (60 mL). After 45 min at -78 ° C, the obtained solution was triturated with saturated NH ₄ Cl, water and warmed to room temperature. The aqueous phase was separated and extracted with ethyl acetate, decolourised with charcoal, filtered through celite and concentrated in vacuo. The resulting foam was purified by flash chromatography (eluent: petrolatum: ethyl acetate 6: 4) to give 4.64 g of the title compound (yield: 38%).

¹ H-NMR (CDCl 3) δ: 8.42 (s, 2H); 7.88-6.80 (m, 3H); 7.77 (d, 1H, J = 3Hz); 7.38 (d, IH); 5.37 (s, 1 H); 3.87 (s, 1 H); 3.83 (s, 3H).

Example 145

3- (3,5-Dichloro-1-oxypyridin-4-ylmethylene) -6-methoxy-1-thiazol-2-yl-1,3-dihydroisobenzofuran-1-ol

Under N ₂ m-chloroperbenzoic acid (4.15 g, 13.2 mmol) was added to a solution of 3- (3,5-dichloropyridin-4-ylmethylene) -6-methoxy-1-thiazol-2-yl-1,3- of dihydroisobenzofuran-1-ol (4.47 g, 11 mmol), prepared as described in Example 144, in CHCl 3 (45 mL) and the mixture was refluxed overnight. Additional m-chloroperbenzoic acid (0.34 g) was added and the mixture was refluxed again, then cooled with ice. The solid was filtered and washed with CHCl ₃ and ethyl ether to give a crude solid. The mother liquors were washed with water, Na ₂ S ₂ O ₅ and water / Na ₂ CO ₃ , decolourised with charcoal, filtered through celite and concentrated in vacuo. The solid obtained was combined with the previous portions and purified by flash chromatography (eluent CH ₂ Cl ₂ : CH ₃ OH 95: 5) to give a solid which was triturated in ethyl ether to give 2.56 g of the title compound (yield: 55%). ). Melting point: 230 - 233 ° C (dec.)

0

- 102 ·· 0 · 00 0 00

0000 000 ·

0 0000 00 0 0 0 0 0 0 0000 · 0 0 ··· 00 »00> 00000 00> 0«>

¹ H-NMR (DMSO) δ: 8.59-8.55 (m, 2H); 7.80-7.71 (m, 2 H); 7.776.82 (m, 3H); 7.33 (s, 1 H); 5.22 (s, 1 H); 3.81 (s, 3H).

Example 146

1- (3,5-Dichloro-1-oxopyridin-4-ylmethyl) -6-methoxy-4-thiazol-2-yl-phthalazine (Compound 85)

Acetic acid (1.68 g, 28 mmol) and hydrazine monohydrate (0.3 g, 5.88 mmol) were added to a suspension of 3- (3,5-dichloro-1-oxy-pyridin-4-ylmethylene) -6 -methoxy-1-thiazol-2-yl-1,3-dihydroisobenzofuran-1-ol (2.36 g, 5.6 mmol), prepared as described in Example 145. The mixture was refluxed for 5 hours , allowed to stand for 2 days, then refluxed overnight. Additional hydrazine monohydrate (5.88 mmol) was added and the mixture was refluxed overnight, cooled with ice and filtered, washed with plenty of water. After drying at 60 ° C under vacuum, the obtained solid was purified by chromatography (eluent: ethyl acetate) to give a solid which was triturated in ethyl ether to give 1.59 g of the title compound. Melting point: 248.4-249.4 ° C (dec.).

¹ H-NMR (CDCl 3) δ: 9.35 (d, 2H, J = 2.6 Hz); 8.22 (s, 2 H); 8.14 (d, 1H, J = 9.1Hz); 8.06 (d, 1H, J = 3.3Hz); 7.61 (dd, IH); 7.51 (d, IH); 4.87 (s, 2 H); 4.07 (s, 3H).

Example 147

PDE 4 enzyme inhibition

a) Isolation of human polymorphonuclear leukocytes

Polymorphonuclear leukocytes (PMN) were isolated from peripheral blood of healthy volunteers by the method of Boyum A. (Scand. J. Immunol., 1976, 5th suppl., 9). Briefly, PMNs were purified • β 9 • 9

- 10399 99

9 9 · 9

9 9

9 9 9 9 9 9999

9 9 9 9 9 • 9 9999 99 9 by Ficoll-Paque gradient centrifugation followed by sedimentation on dextran, and erythrocyte contamination was removed by lysis in a hypotonic environment.

b) Purification of PDE 4

Human PMNs were suspended in TRIS / HCl buffer (10 mM, pH 7.8) containing MgCl ₂ (5 mM), EGTA (4 mM), mercaptoethanol (5 mM), TRITON-X100 (1%), pepstatin A ( 1 μΜ), PMSF (100 μΜ) and leupeptin (1 μΜ), and homogenized on a Polytron homogenizer. The homogenate was centrifuged at 25,000 xg for 30 min at 4 ° C and PDE 4 was purified by ion exchange chromatography using the FPLC technique of Schudt C. et al. (Naunyn-Schmidberg's Arch. Pharmacol., 1991, 334, 682). loaded onto a UNO Q12 column (Bio-Rad) and the enzyme eluted with a linear gradient of sodium acetate from 50 mM to 1 M at a flow rate of 4.5 mL / min. Fractions containing enzymatic activity were pooled, dialyzed against water and concentrated. The PDE 4 enzyme was stored at -20 ° C in the presence of ethylene glycol (30% v / v) until use.

(c) PDE 4 activity test

Enzymatic activity was evaluated using a Scintillation Proximity Assay (SPA) kit (Amersham). The enzymatic reaction was performed in a final volume of 100 μί TRIS / HCl buffer (50 mM, pH 7.5), MgCl ₂ (8.3 mM), EGTA (1.7 mM), cAMP (1 μ 1) and [ ³ H] cAMP (~ 100,000 dpm) as a tracer. Compounds of the invention, reference compounds, or vehicle were added at various concentrations. 6,7-Dimethoxy-4- (pyridin-4-ylmethyl) -2H-phthalazin-1-one (reference 1) and 6,7-dimethoxy-4- (piperidin-4-yl-) were used as reference compounds. methyl) -2 / 7-phthalazin-1-one (ref. 2), which are included in the general formula of EP-0 722 936 (Eisai). The reaction was initiated by the addition of 1.5 µg protein and the mixture was incubated for 40 min at 30 ° C. SPA beads (50 μί) containing 18 mM zinc sulfate were added to stop the reaction and after 20 min at temperature

- 104 99 99 • 9 * 9 9

9 9 9

9 9 • 9 9999

9

9 9

9 9 9

9 9999

9 9

9 ‘99 of the laboratory, radioactivity was measured using a scintillation counter.

The IC ₅₀ value refers to the nanomolar concentration of the compound required to inhibit the cyclic nucleotide hydrolysis by 50%, and was calculated by non-linear regression analysis.

The compounds of formula I of the present invention are capable of selectively inhibiting PDE 4. The results are shown in Table 1 below.

Table 1

Compound IC50 nM 5 37 ± 6 8 51 ± 7 9 36 ± 4 10 28 ± 4 11 26 ± 3 12 30 ± 8 13 12 + 2 15 Dec 21 ± 4 16 72 ± 16 17 131 ± 24 20 May 158 ± 34 22nd 225 ± 35 23 209 ± 26 24 36 + 9 26 14 ± 2 27 Mar: 48 + 12 29 241 ± 73

- 105 Table 1 - continued

Compound IC ₅₀ nM 32 10 ± 2 33 19 ± 3 35 7.1 ± 0.5 36 96.4 ± 7.3 37 61 ± 2.4 38 132 ± 30 55 312 ± 48 56 364 ± 67 60 93 ± 29 Reference 1 > 100 μΜ Reference 3 > 100 μΜ

Example 148

Inhibition of TNF release

a) Isolation of human monocytes

Monocytes were isolated from peripheral blood of healthy volunteers according to the procedure of Schreek L. et al. (J. Natl. Cancer Inst., 1964, 32, 507). The monocyte and lymphocyte population was isolated by gradient centrifugation on Ficoll, and cells diluted to a density of 2.5 χ 10 ⁶ cells / ml in RPMI1640 incubation medium containing 1% heat-inactivated fetal calf serum, penicillin (100 U / ml) and streptomycin (100 pg / ml) were plated in 24-well plates (1 ml / well) and allowed to attach for 1 hour at 37 ° C at 5% CO ₂ . Non-adherent lymphocytes were removed by aspiration and the monocytes attached to the plate were used in the next step.

- 106 -

(b) TNF release test '

TNF release _from human monocytes was measured according to the authors of Barnette M. et al (Biochemical Pharmacology, 1996; 51, 949). Monocytes were incubated for 1 hour with 1 ml of RPMI1640 incubation medium (1% heat-inactivated fetal calf serum, 100 U / ml penicillin and 100 µg / ml streptomycin) containing various concentrations of the products of the present invention or vehicle as controls. TNFα _and monocyte release was induced by the addition of 1 ng / ml LPS (E. coli lipopolysaccharide) and after 16 hours incubation at 37 ° C with 5% CO 2, the incubation medium was removed, centrifuged and the supernatant was stored at -80 ° C until run. TNF assay _and ELISA kit (Amersham). Results are expressed as IC ₅₀ values obtained by the same calculation as explained in Example 147.

Table 2

Compound IC50 nM 5 46 ± 15 8 48 ± 14 29 73 ± 21 38 62 ± 17 49 9.1 ± 2.9 ⁵⁶ 260 ± 44

AND

Example 149

Inhibition of PDE 3 and PDE 5 enzymes

a) Preparation of human platelets

Human platelets were prepared from platelet rich plasma (PRP) obtained from healthy volunteers. PRP

- 107 -

was centrifuged at 2200 rpm for 15 min at 4 ° C and the plates were suspended in lysis solution (15 mL; 155 mM NH ₄ Cl, 10 mM KHCO 3 and 0.1 mM Na 2 EDTA, pH = 7.4) and incubated for 10 min in ice bath to remove contaminating erythrocytes. After centrifugation at 1400 rpm for 10 min at 4 ° C, the plates were suspended in 10 ml of 145 nM NaCl, 5 mM KCl, 1 mM MgSO ₄ , 10 mM glucose, 10 mM HEPES (pH 7.4) and 0.05 U / ml hirudin, and stored at -20 ° C until homogenization. The plates were thawed and 50 ml of 20 mM TRIS (pH = 6.5) containing 5 mM β-mercaptoethanol, 2 mM | EDTA, 50 mM sodium acetate and 50 μΜ PMSF (homogenization buffer).

The plate suspension was then homogenized on a Polytron (Polytron PT 1200) homogenizer for 20 sec. The homogenate was centrifuged at 14,500 rpm for 20 min at 4 ° C, and the supernatant was applied to a UNO Q12 column (Bio-Rad). The enzymes PDE 3 and PDE 5 were eluted with a linear gradient of sodium acetate from 0.05M to 1M at a flow rate of 4.5 ml / min. Fractions containing enzymatic activities (PDE 3 or PDE 5) were pooled, dialyzed against water and concentrated ten times by ultrafiltration. The PDE 3 and PDE 5 fractions were stored at -20 ° C in the presence of ethylene glycol (30% v / v) until use.

(b) PDE 3 and PDE 5 activity assay

Enzymatic activity was evaluated by the Scintillation Proximity Assay (SPA) kit (Amersham). The enzymatic reaction was performed in a final volume of 100 μΙ TRIS / HCl buffer (50 mM, pH 7.5), MgCl ₂ (8.3 mM), EGTA (1.7 mM), cAMP (for PDE 3 assay) or cGMP (for assay). PDE 5) (1 μΜ), [ ³ H] cAMP or [ ³ H] cGMP (10 μΙ), and

I 10 μΙ of compounds of the invention or vehicle. The reaction was initiated by addition of enzyme (1.0 µg) and incubation for 40 min at 30 ° C.

SPA beads (50 μΙ) containing 18 mM zinc sulfate were added to stop the reaction, and after 20 min at room temperature, radioactivity was measured using a scintillation counter.

Compounds were tested at a concentration of 10 ^-6 M and the results are

- 108 expressed as percent inhibition.

Table 3

Compound % PDE 3 inhibition at 10 ^-6 M % PDE 5 inhibition at 10 ^-6 M 5 7 21 8 12 35 11 17 10 15 Dec 15 Dec 0 17 12 21 20 May 5 11 22nd 20 May 8 23 19 Dec 21 26 - 0 27 Mar: 15 Dec 9 29 -4 1 38 -2 1 49 -10 10 54 2 33 56 -9 14 60 -2 1

Represented by:

• · · · · ·

Claims (11)

What is claimed is: 1. A compound of formula (I): EMI3.0 wherein - is a single or double bond; Z is NH, methylene, (C ₂ -C ₆₎ alkylene chain which is optionally branched and / or unsaturated and / or interrupted by a (C5-C7) cycloalkyl radical; A is phenyl or heterocycle optionally substituted with one or more substituents selected from oxo, nitro, carboxy and halogen, or COR ₄ wherein R ₄ is hydroxy, (C 1 -C 6 alkoxy) amino optionally substituted with one or two (C 1 -C 6) alkyl hydroxy groups; R is (C 1 -C 6) alkyl or polyfluoro (C 1 -C 6) alkyl a group; R 1 is absent if - represents a double bond or, if - represents a single bond, represents a) a hydrogen atom; • 9 9 9 9 9 9 9 9999 9 9 9 9 9 99 9999 9 Π-1Π 999 99 99 - I JU - 99 9900 99 99 99 9 b) (C 1 -C ₆ ) alkyl optionally substituted with aryl, heterocycle or COR ₅ wherein R ₅ is hydroxy, (C 1 -C ₄ ) alkoxy or hydroxyamino; c) -COR ₆ wherein R ₆ is hydrogen, aryl, aryl- (C 1 -C ₆ ) alkyl, amino optionally alkylated or monohydroxylated, hydroxy, (C 1 -C ₄ ) alkoxy, carboxy, (C 1 -C ₄ ) alkoxycarbonyl, HN = C-NH ₂ , or (C 1 -C 4 alkyl optionally substituted with a heterocycle; d) (C 1 -C ₄ ) alkylsulfonyl; R ₂ represents two hydrogen atoms or the group = 0, where - is a single bond, or when - is a double bond, R ₂ is hydrogen, cyano, _(Ci-4) alkoxycarbonyl, amido, optionally substituted aryl or heterocycle, (C C ₈ ) alkyl, (C ₂ -C ₈ ) alkenyl or (C ₂ -C ₈ ) alkynyl optionally branched and / or substituted with aryl or heterocycle; aryloxy, heterocyclyloxy, aryl- (C 1 -C ₄ ) -alkoxy, heterocyclyl- (C 1 -C ₄ ) alkoxy, amino substituted with one or two (C 1 -C ₄ ) alkyl groups, arylamino, heterocyclylamino, aryl- (C 1 -C ₄ ) alkylamino , heterocyclyl _icv-C4) alkylamino; R ₃ is hydrogen, or a (C 1 -C ₈ ) alkyl, (C ₂ -C ₈ ) alkenyl or (C ₂ -C ₈ ) alkynyl optionally substituted with hydroxy, oxo, aryl or heterocycle, and optionally interrupted by one or more heteroatoms or one or more hetero groups; N -> O derivatives of compounds of formula I and pharmaceutically acceptable salts thereof; provided that R ₂ is pyridyl, R is methyl, R is absent and R3 is hydrogen and Z is not NH. A compound according to claim 1, wherein - represents a double or single bond; and Z is methylene or a _(C2-C6) alkylene chain. -111 The compound of claim 1, wherein R 22 is double or single; custody; Z is methylene or a _(C2 -CE) alkylene chain; and A represents a heterocycle optionally substituted with one or more substituents. The compound of claim 1, wherein - represents a double or single bond; Z is methylene; and A is pyridine substituted with two substituents. The compound of claim 5, wherein - is a single bond and R ₂ is two hydrogen atoms. The compound of claim 5, wherein - represents a double bond; and R ₂ is cyano, (C 1 -C 4) alkoxycarbonyl, amido, optionally substituted heterocycle, (C ₂ -C 8) alkenyl or (C ₂ -C 8) alkynyl optionally substituted with aryl or heterocycle; aryloxy, heterocyclyloxy, arylamino, heterocyclylamino. A compound according to claim 7 of the formula AND · 9 wherein R ₂ is a heterocycle. AND The compound of claim 1 which is: 1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-4-phenyl-phthalazine; 4- (3,5-dichloropyridin-4-ylmethyl) -7-methoxy-1H-phthalazine-2-carboxylic acid methyl ester; benzyl- {3- [1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl] -prop-2-ynyl} -methylamine; 1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-5- (5-morpholin-4-yl-pent-1-ynyl) -phthalazine dihydrochloride; 3- [1- (3,5-dichloro-pyridin-4-ylmethyl) -6-methoxy-phthalazin-5-yl] -prop-2-yn-1-ol; 1- (3,5-Dichloro-pyridin-4-ylmethyl) -6-methoxy-4-morpholin-4-yl-phthalazine; ‘1- (3,5-dichloropyridin-4-ylmethyl) -6-methoxy-4- [1,2,4] triazol-1-yl-phthalazine. A pharmaceutical composition comprising a therapeutically effective amount according to claim 1 together with a suitable carrier. Pharmaceutical composition according to claim 10 for allergic and inflammatory diseases. healing - 113 44 44 444 9 4 4 4 4 4 5 4 4 4 4 4 4 4 4 44 44444 »4444 Pharmaceutical composition according to claim 10 for respiratory diseases. healing Represented by: